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[post_date] => 2024-11-15 15:04:30
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ZipChip CE-MS was employed to assess mAb quality attributes directly from spent media, requiring only simple dilution without additional preparation. This technique facilitated daily monitoring of molecular weight, charge, and glycoform heterogeneity in mAbs produced in bioreactors with different glucose feeding strategies. Results showed improved product quality attributes from continuous glucose feeding compared to those produced with bolus glucose feeding, demonstrating ZipChip’s potential as a simple and effective tool for real-time monitoring of product quality in bioprocess development.
[post_title] => Enhanced Bioprocess Control: Assessing mAb Quality Through Direct Spent Media Analysis with ZipChip CE-MS
[post_excerpt] => ZipChip CE-MS charge variant analysis enables streamlined monitoring of mAb quality attributes throughout cell culture, supporting bioprocess development.
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ZipChip CE-MS was used to analyze biosimilar mAb candidates, resolving charge variants and glycoforms with high sensitivity and short run times. This approach enabled rapid characterization of critical quality attributes, such as glycosylation and charge modifications, from early-stage cell line candidates. By providing detailed insights into post-translational modifications in a single assay, ZipChip facilitated efficient selection of mAbs closely matching the originator's quality profile, accelerating biosimilar development.
[post_title] => Sensitive Screening Of Biosimilar Candidate mAbs From Clonal Cell Lines Using ZipChip CE-MS
[post_excerpt] => ZipChip CE-MS enables rapid and sensitive analysis of mAb glycosylation profiles and charge variants, streamlining biosimilar candidate selection from early-stage clone screening.
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[post_date] => 2023-05-01 11:52:32
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[post_title] => Microchip CE/MS Peptide Mapping Workflow with Trypsin Digestion for the Analysis of Critical Quality Attributes of Monoclonal Antibodies
[post_excerpt] => Peptide mapping is considered the gold standard for the analysis of biopharmaceuticals, including monoclonal antibodies.
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[post_title] => Rapid characterization of oligonucleotides using microfluidic CE-MS by ZipChip
[post_excerpt] => This application note presents a CE-MS analysis of oligos by a ZipChip coupled to a QExactive HF mass spectrometer. This positive ESI mode method utilizes a MS-friendly background electrolyte (BGE), that eliminates the use of ion-paring agents such as hexafluoro isopropanol or alkyl amines.
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[post_title] => Charge variant analysis with ZipChip and maXisII
[post_excerpt] => A ZipChip capillary zone electrophoresis system coupled to Bruker maXis II UHR-QTOF offers an alternative selectivity to separate charge variants prior to MS analysis and affords a more complete intact mass characterization without adding complexity or high salt eluents.
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[post_date] => 2021-09-10 13:33:28
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[post_title] => A ZipChip Based CZE-MS Analysis for Quick Assessment of Biotherapeutics
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[post_date] => 2021-06-21 14:28:37
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[post_title] => From comprehensive characterization to monitoring product degradation characteristics
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[post_name] => zipchip-native-charge-variant-analysis-of-biotherapeutics-using-zipchip-ce-esi-ms
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[post_date] => 2020-12-11 19:14:54
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[post_title] => Charge Variant Analysis Coupled to a SCIEX TripleTOF® 6600+ System Mass Spectrometer
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[post_date] => 2020-11-16 23:18:26
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[post_title] => High-Throughput Intact Mass Analysis of mAb Based Therapeutics
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[post_title] => Analysis of Reduced mAbs and ADCs
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[post_name] => analysis-reduced-mabs-antibody-drug-conjugates-using-zipchip-platform
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[post_title] => ZipChip for profiling and characterization of antibody charge heterogeneity
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[post_title] => Sciex Flexible Solution for MAM with ZipChip
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[post_name] => zipchip_app-note_sciex-charge-variant-liability-analysis-using-the-sciex-flexible-solution-for-mam
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[post_title] => Sciex Presents Charge Heterogeneity of Monoclonal Antibodies with ZipChip
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[post_title] => Charge Variant Analysis of Native Antibodies for In-depth Characterization of mAb-based Biotherapeutics
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ZipChip CE-MS was employed to assess mAb quality attributes directly from spent media, requiring only simple dilution without additional preparation. This technique facilitated daily monitoring of molecular weight, charge, and glycoform heterogeneity in mAbs produced in bioreactors with different glucose feeding strategies. Results showed improved product quality attributes from continuous glucose feeding compared to those produced with bolus glucose feeding, demonstrating ZipChip’s potential as a simple and effective tool for real-time monitoring of product quality in bioprocess development.
[post_title] => Enhanced Bioprocess Control: Assessing mAb Quality Through Direct Spent Media Analysis with ZipChip CE-MS
[post_excerpt] => ZipChip CE-MS charge variant analysis enables streamlined monitoring of mAb quality attributes throughout cell culture, supporting bioprocess development.
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Application Notes & Snapshots
Enhanced Bioprocess Control: Assessing mAb Quality Through Direct Spent Media Analysis with ZipChip CE-MS
ZipChip CE-MS charge variant analysis enables streamlined monitoring of mAb quality attributes throughout cell culture, supporting bioprocess development.
Sensitive Screening Of Biosimilar Candidate mAbs From Clonal Cell Lines Using ZipChip CE-MS
ZipChip CE-MS enables rapid and sensitive analysis of mAb glycosylation profiles and charge variants, streamlining biosimilar candidate selection from early-stage clone screening.
Microchip CE/MS Peptide Mapping Workflow with Trypsin Digestion for the Analysis of Critical Quality Attributes of Monoclonal Antibodies
Peptide mapping is considered the gold standard for the analysis of biopharmaceuticals, including monoclonal antibodies.
Rapid characterization of oligonucleotides using microfluidic CE-MS by ZipChip
This application note presents a CE-MS analysis of oligos by a ZipChip coupled to a QExactive HF mass spectrometer. This positive ESI mode method utilizes a MS-friendly background electrolyte (BGE), that eliminates the use of ion-paring agents such as hexafluoro isopropanol or alkyl amines.
Charge variant analysis with ZipChip and maXisII
A ZipChip capillary zone electrophoresis system coupled to Bruker maXis II UHR-QTOF offers an alternative selectivity to separate charge variants prior to MS analysis and affords a more complete intact mass characterization without adding complexity or high salt eluents.
A ZipChip Based CZE-MS Analysis for Quick Assessment of Biotherapeutics
From comprehensive characterization to monitoring product degradation characteristics
Charge Variant Analysis Coupled to a SCIEX TripleTOF® 6600+ System Mass Spectrometer
High-Throughput Intact Mass Analysis of mAb Based Therapeutics
Analysis of Reduced mAbs and ADCs
ZipChip for profiling and characterization of antibody charge heterogeneity
Sciex Flexible Solution for MAM with ZipChip
Sciex Presents Charge Heterogeneity of Monoclonal Antibodies with ZipChip
Charge Variant Analysis of Native Antibodies for In-depth Characterization of mAb-based Biotherapeutics
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Our ZipChip CVA kit takes the guesswork out of method development. Characterizing your samples with ZipChip capillary electrophoresis coupled to your high-resolution MS, you're able to assess charge heterogeneity, identify basic and acidic variants, acquire accurate mass information, and profile glycoforms, all in a single analysis.
Get more data and streamline your Charge Variant Analysis workflow with ZipChip CVA kit
- Easy sample prep and rapid analysis
- Little-to-no method development
- High performance analysis with full charge variant separation and peak identification
- Comparable profiles to commonly used CVA techniques with the added power of mass spec characterization
- Dilute Sample and Prepare BGE
- Load Sample into ZipChip
- Run the CVA on ZipChip-MS
Get straight to it.
Our preconfigured ZipChip CVA kit takes the guesswork out of method development.
SEAMLESS WORKFLOW
ZipChip-MS peak profile is comparable to Capillary Isoelectric Focusing (CiEF), Cation Exchange Chromatography (CEX), and Capillary Zone Electrophoresis (CZE-UV) and delivers seamless workflow for identification of detected unknown variant peaks.
STREAMLINED DESIGN
Analyze multiple biotherapeutic protein CQAs in a single assay - gain the ability to clearly separate and identify acidic and basic species and assess the glycosylation profile.
OPTIMIZED FOR YOU
Characterize a variety of monoclonal antibodies (mAbs) and other biotherapeutic proteins with the ZipChip CVA kit without having to reoptimize your method.
ZipChip CVA Consumables
ZipChip CVA consumables provide everything you need for analysis with no method development and minimal sample prep.
Name Contents Sample per kit Part No. CVA Assay Kit CVA Background Electrolyte (BGE), Diluent, and Additive 625 850-00052 HRN Chip (5 pack) High Resolution Native Chip (HRN) x5 625 810-00227
[post_title] => Full Charge Variant Analysis with Peak Identification Using ZipChip CE-MS
[post_excerpt] => ZipChip CVA kit.
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ZipChip
CONSUMABLES GUIDE
From complex biologics to small molecules,
ZipChip has an assay kit for you.
Take the Guesswork Out of Method Development
3 Easy Steps to Selection
1. Application
2. Chip
3. Kit
Assay kits come with background electrolyte and sample diluent that’s optimized for each application. Just put the bottle of background electrolyte in the autosampler, prep your samples with the sample diluent and you’re off.
1. The Application
From large to small molecule, ZipChip’s got you covered.
Protein Characterization:
ZipChip is a one-stop shop for CQA’s—from charge variant analysis to peptide mapping. You’ll get high-resolution separation coupled to MS with simple, streamlined workflows.
Metabolomics: Quickly analyze small polar analytes from a variety of matrices, or monitor dynamic levels of metabolites in real time. Sample prep is quick and easy and results are ready in minutes.
Oligonucleotides: The characterization of nucleic acid-based drugs like oligonucleotides isn’t easy. ZipChip can offer some advantage over LC separations, particularly variations due to charge or shape.
Small Molecules: Analysis is easy and fast with ZipChip. Simply dilute and shoot; there’s no labeling or derivatization. Small polar analyte assays take as little as 2 minutes. Add internal standards for full quantitation on a variety of analytes.
2. The Chips
Choose from a range of chip options to optimize your workflow. Each chip can be used up to 125 injections.
HS: High Speed chip enables fastest separations. Matching BGE kits: Intact Antibody, Peptides, or Metabolites.
HR: High Resolution chip for applications demanding maximum separation capacities. Matching BGE kits: Intact Antibody, Peptides, or Metabolites.
HSN: High Speed chip for fast native protein analysis. Matching BGE kits: Native Antibodies or Charge Variant Analysis.
HRN: High Resolution chip with ultimate separation for native protein analysis. Matching BGE kits: Native Antibodies, or Charge Variant Analysis.
HSB: High Speed Bare Glass chip for high throughput separations. For example: fast oligonucleotides analysis. Matching BGE kit example: Oligos BGE.
HRB: High Resolution Bare Glass chip for high resolution separation of negatively charged analytes such as oligonucleotides. Matching BGE kit example: Oligos BGE.
3. BGE Kits
Background electrolyte (BGE) and sample diluent reduce preparation time for your samples. With ZipChip you just dilute and shoot!
Charge Variant Analysis: Formulated to provide excellent charge variant separations and MS sensitivity for antibodies and other proteins on all mass spectrometers.
Native Antibodies: Excellent charge variant separation of antibodies and other constructs to generate native spectra.
Peptides: Fast, efficient separations of protein digests and peptides utilizing low pH.
Metabolites: Streamlined sample prep and high-speed analysis for small molecules in denaturing conditions at low pH.
Oligonucleotides: Rapid analysis of oligonucleotides without the use of ion-pairing agents.
Ordering Information
ZipChip Assay Kits* Part Number Samples Per Kit Charge Variant Analysis Kit 850-00052 625 Native Antibody Assay 850-00048 625 Metabolites Assat 850-00033 625 Peptide Assay 850-00034 625 Oligos Assay 850-00066 625 Oligos Starter 850-00067 250 Tests
*All kits contain BGE and diluent, and chips need to be ordered separately. The exception is the oligos starter kit which contains BGE, Diluent, chips and a standard.
Chips Part Number Chips Per Box Injections Per Chip HS 810-00195 5 125 HR 810-00194 5 125 HSN 810-00230 5 125 HRN 810-00227 5 125 HSB 810-00237 5 125 HRB 810-00239 5 125
[post_title] => ZipChip Consumables Guide
[post_excerpt] => Learn how to take the guesswork out of method development with ZipChip. Breeze through your selection in three quick steps.
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HASSLE FREE
Oligo characterization with ZipChip
The newly released ZipChip® oligos kit provides a simple, sensitive, dilute-and-shoot method for rapid analysis of oligonucleotides, applicable to various sizes and modifications.
Benefits:
- High-resolution accurate mass identification and quantitation
with CE-MS
- No ion-pairing agents; no instrument contamination
- Orthogonal to LC-based separation
- Analysis in minutes with simple sample prep
Part Name Contents Samples per kit Part No. ZipChip Oligos Starter Kit Oligos Background electrolyte (BGE) x 2
Diluent x 2
HRB Chip x 2
Oligo standard 250 850-00067 ZipChip Oligos Kit Oligos BGE x 5
Diluent x 5 625 850-00066 HSB Chip High Speed Bare Glass Chip (HSB) x 5 625 810-00237 HRB Chip High Resolution Bare Glass Chip (HRB) x 5 625 810-00239 ZipChip Oligos Standard 20 mer DNA Primer - 445-02281
Rapid Characterization of Oligos
Separation of Modified ssRNAs
Electropherogram showing separation of biotinylated, glycosylated and phosphorylated 20 mers from the corresponding unmodified 20 mer RNA standard
Rapid characterization of ssRNA with no method development
Base Peak Electropherogram for 20mer unmodified ssRNA standards (left) along with the raw mass spectrum (right).
[post_title] => Hassle-Free Oligo Characterization with ZipChip
[post_excerpt] => The newly released ZipChip® oligos kit provides a simple, sensitive, dilute-and-shoot method for rapid analysis of oligonucleotides, applicable to
various sizes and modifications.
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ZipChip leverages microfluidic-based Capillary Electrophoresis (CE) and electrospray ionization technology to simplify sample preparation, perform high-resolution separations with reduced analysis times, and directly introduce samples into a Thermo Fisher Scientific, Bruker, or Sciex mass spectrometer. Achieve faster results with the ZipChip CE-MS device.
CE-MS DEVICE
ZipChip
ACCELERATE YOUR MS CHARACTERIZATION
Rapid protein characterization and metabolomic analysis with microfluidic technology for highly efficient separations of complex samples.
BIOLOGICAL CHARACTERIZATION
IN MINUTES
GIVE YOUR MASS SPEC SOME ZIP
The ZipChip® platform prepares and separates a wide range of biological samples, then electrosprays them into your mass spec for analysis. Just clip it on. The process takes as little as a few minutes, and it results in better separation quality than most LCs in a fraction of the time. Simple workflows and multiple kit options cover a host of biotherapeutic, metabolomic, and proteomic applications.
EASY ON SAMPLES
Now the kicker. ZipChip plays well with your mass spec and your proteins. Gentle sample prep reagents preserve structural integrity without denaturing or unfolding proteins for fully native mass spectra. No need to remove detergents or desalt. Negative and neutrals are trashed. Only positive analytes head out for cleaner mass spectra and more identified peaks.
EFFICIENCY WITH ADVANCED
MICROCHIP TECHNOLOGY
WITH ZIPCHIP YOU GET THE DATA FASTER, AND BECAUSE OF THAT YOU CAN MAKE QUICK DECISIONS
Associate scientist at Amgen
Easy as 1-2-3
Pick an application, a chip, and a kit. After simply loading the premixed background electrolyte and sample into the autosampler, pop in a ZipChip, and hit run. Use vials or 96-well plates.
It’s hands-off from there. Smart chips tell the system which method to use based on your preferences. Samples are automatically loaded, separated, and electrosprayed. Integrated software cues the mass spec to start analysis.
Efficient Separations
Microfluidic technology integrates capillary electrophoresis (CE) and electrospray ionization (ESI) on the ZipChip. Junctions and dead volumes don’t exist so analytical quality stays high, and no injection bias improves repeatability.
1. Pick your application, kit, and chip
2. Simple sample prep
3. Load reagent and chip, then run
IT'S ALL IN THE CHIP
FROM LARGE TO SMALL MOLECULES,
ZIPCHIP’S GOT YOU COVERED
Protein Characterization:
ZipChip is a one-stop shop for CQAs—with streamlined CE MS workflows for Charge Variant Analysis of basic and acidic species, Rapid Intact Mass Analysis, Subunit Analysis, and Peptide Mapping for deeper characterization.
Oligonucleotides:
The characterization of nucleic acid-based drugs like oligonucleotides isn’t easy. ZipChip can offer some advantage over LC separations, such as no ion-pairing agents to contaminate the instrument
Small Molecules
Analysis is easy and fast with ZipChip. Simply dilute and shoot; there’s no labeling or derivatization. Small polar analyte assays take as little as 2 minutes. Add internal standards for full quantitation on a variety of analytes.
Metabolomics:
Detect, identify and quantitate small polar analytes from a variety of matrices like cell lysates or growth media and monitor dynamic levels of metabolites in real time. Sample prep is quick and easy and results are ready in minutes.
[post_title] => ZipChip: Accelerate Your MS Characterization
[post_excerpt] => Rapid protein characterization and metabolomic analysis with microfluidic technology for highly efficient separations of complex samples.
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Our ZipChip CVA kit takes the guesswork out of method development. Characterizing your samples with ZipChip capillary electrophoresis coupled to your high-resolution MS, you're able to assess charge heterogeneity, identify basic and acidic variants, acquire accurate mass information, and profile glycoforms, all in a single analysis.
Get more data and streamline your Charge Variant Analysis workflow with ZipChip CVA kit
- Easy sample prep and rapid analysis
- Little-to-no method development
- High performance analysis with full charge variant separation and peak identification
- Comparable profiles to commonly used CVA techniques with the added power of mass spec characterization
- Dilute Sample and Prepare BGE
- Load Sample into ZipChip
- Run the CVA on ZipChip-MS
Get straight to it.
Our preconfigured ZipChip CVA kit takes the guesswork out of method development.
SEAMLESS WORKFLOW
ZipChip-MS peak profile is comparable to Capillary Isoelectric Focusing (CiEF), Cation Exchange Chromatography (CEX), and Capillary Zone Electrophoresis (CZE-UV) and delivers seamless workflow for identification of detected unknown variant peaks.
STREAMLINED DESIGN
Analyze multiple biotherapeutic protein CQAs in a single assay - gain the ability to clearly separate and identify acidic and basic species and assess the glycosylation profile.
OPTIMIZED FOR YOU
Characterize a variety of monoclonal antibodies (mAbs) and other biotherapeutic proteins with the ZipChip CVA kit without having to reoptimize your method.
ZipChip CVA Consumables
ZipChip CVA consumables provide everything you need for analysis with no method development and minimal sample prep.
Name Contents Sample per kit Part No. CVA Assay Kit CVA Background Electrolyte (BGE), Diluent, and Additive 625 850-00052 HRN Chip (5 pack) High Resolution Native Chip (HRN) x5 625 810-00227
[post_title] => Full Charge Variant Analysis with Peak Identification Using ZipChip CE-MS
[post_excerpt] => ZipChip CVA kit.
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Brochures & Flyers
Full Charge Variant Analysis with Peak Identification Using ZipChip CE-MS
ZipChip CVA kit.
ZipChip Consumables Guide
Learn how to take the guesswork out of method development with ZipChip. Breeze through your selection in three quick steps.
Hassle-Free Oligo Characterization with ZipChip
The newly released ZipChip® oligos kit provides a simple, sensitive, dilute-and-shoot method for rapid analysis of oligonucleotides, applicable to
various sizes and modifications.
ZipChip: Accelerate Your MS Characterization
Rapid protein characterization and metabolomic analysis with microfluidic technology for highly efficient separations of complex samples.
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Getting life-changing biologics to patients ASAP
HJB’s goal is to overcome scientific and technical hurdles that inhibit patients’ access to life-changing biologics. To do that, they aim to provide high quality, holistic, and reliable services to innovative biopharma companies.
So how exactly do they hit that lofty goal? The contract development and manufacturing organization (CDMO) based in Zhejiang, China, employs cutting-edge technology that substantially accelerates R&D processes, reduces costs, and fast-tracks commercialization. They offer biopharma clients access to complete, flexible, and reliable development and manufacturing services that include cell line development, bioprocess development, analytical method development, formulation development and GMP manufacturing for all clinical phases and commercial production.
Overcoming hurdles in cell culture media analysis
CHALLENGE: The waiting game
Prior to HJB’s integration of the REBEL™ cell culture media analyzer into their processes, the upstream process development team, led by Vice President Lisa Zheng, sent media samples off to HJB’s analytical sciences group for amino acid analysis. But relying on another team for results meant they were waiting to make decisions and take next steps in their media development processes.
In their search to accelerate R&D and reduce costs, HJB discovered the power of REBEL and ZipChip.
SOLUTION: Answers at-line, anytime
Lisa’s team purchased a REBEL and set it up right next to their bioreactors. They found the device space-friendly and easy to install. Consumables were easy to maintain, and operation was so simple that even their non-technical analysts could conduct analysis. Samples were measured quickly, and with results in less than 10 minutes, scientists were making decisions in near-real time. REBEL is now used routinely for quick analysis to support accelerated media and process improvements.
CONCLUSION: A new milestone in productivity
Because REBEL lets the upstream process development group analyze many samples in a short period of time, the group accumulates a large amount of amino acid data just through daily testing. This lets them quickly investigate deviations during product development, carry out process development/optimization based on REBEL data, and better understand the impacts on culture performance and Product Quality Attributes (PQAs) from process parameter changes.
REBEL was also key in helping HJB accelerate media and process development, which is essential for the development of highly intensified, continuous cell culture perfusion. Earlier this year, the company announced that its continuous perfusion cell culture platform has achieved industry-leading volumetric productivity of greater than 6 g/L per day while maintaining process and product quality attributes in a state of control for a 4-week culture.
“REBEL can analyze a large number of samples in a short period of time to provide valuable data to accelerate cell culture process development. Rather than obtaining amino acid profiles after the run is completed, REBEL provides real-time data for us to make process decisions while the experiments are in progress to maximize successful outcomes.”
-Dr. Christopher Hwang, CTO of HJB
Overcoming hurdles in cell culture media analysis
CHALLENGE: A fair share of challenges with LC
Liming Shi is the VP in charge of the Analytical Science and Quality Control Departments, and responsible for Product & Process Development Operation. HJB’s Analytical Science Department develops and optimizes analytical methods, and provides analytical support for cell lines upstream, downstream, and formulation process development. The group applies the most advanced analytical techniques to perform large molecule characterization, including analysis of mAbs, ADCs, and bispecific antibodies.
Liming’s team was looking for difficult-to-separate charge variants, which is a challenge with LC-based separations. They were also bumping into UHPLC challenges, such as poor separation and analysis with limited sample volumes. Furthermore, their UHPLC-MS separation and analysis had a turn-around time that was too long.
SOLUTION: In comes ZipChip
In their search for analytical tools that could provide higher-quality data and get them to results faster, the Analytical Services team put ZipChip® straight to work.
Because ZipChip enables rapid measurement of Critical Quality Attributes (CQAs) of native and intact monoclonal antibodies (mAbs), the team used it to characterize intact mAbs and assess the root cause of increases in acidic and basic variants under stress at high temperature. The antibody was analyzed at multiple levels, including its intact state under native conditions and subunit and peptide levels. They used the same ZipChip system and method for both analyses, and were able to determine charge heterogeneity speciation and identification in under 10 minutes. This new workflow was 100 times faster with less prep compared to conventional LC, which required them to harvest fractions.
It was also easy to start integrating ZipChip into their other laboratory workflows as minimal method development was needed for different types of proteins. The team noted this was a big benefit when it came to analysis of native proteins.
Liming Shi -
VP of Analytical Science and Quality Control Departments, Product & Process Development Operation
“ZipChip is a capable platform for multiple CQA assays, and for charge variant analysis it is very straightforward to get separation and identification information with minimal method development needed for different types of proteins, especially for the analysis in native conditions. The sample preparation is simple and straightforward, and the sample requirements are at nanogram levels.”
CONCLUSION: ZipChip + MS goes mainstream
The Analytical Science and Quality Control groups at HJB now have ZipChip systems coupled to their Q Exactive and other Thermo Fisher Scientific mass specs. They’ve adopted ZipChip into their intact antibody analysis workflows (denatured and native conditions) and their subunit and peptide mapping analysis as it gives them very clear and in-depth information on antibody molecules. One of the stand-out features in Liming’s opinion is having an online platform that provides straightforward separation and identification of charge variants based on different mechanisms.
The Analytical Science team recently published their work using ZipChip in Analytical Biochemistry, Volume 625, 15 July 2021, 114214: Characterization and monitoring of charge variants of a recombinant monoclonal antibody using microfluidic capillary electrophoresis-mass spectrometry.
[post_title] => REBEL + ZipChip help HJB Overcome Scientific Hurdles and Optimize Processes in Biotherapeutic Development
[post_excerpt] => In their search to accelerate R&D and reduce costs, HJB discovered the power of REBEL and ZipChip.
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Researchers at the Dana-Farber Cancer Institute and Harvard Medical School leveraged the ZipChip® in a combined structural biology-phosphoproteomic study to decipher key topological features and post-translational modifications on the kinase BRAF. Like most cellular proteins, BRAF does not work alone, but rather assembles with other proteins into ‘molecular machines’ which work in concert to orchestrate the cellular ‘circuitry’ required for normal physiology. However, genetic mutation can commandeer and re-direct the activity of these protein machines leading to human tumors. In fact, BRAF mutations contribute to a large number of different human cancers.
The rapid-fire, 10-minute injection-to-injection throughput provided by the ZipChip enabled the researchers to analyze hundreds of samples spanning different BRAF mutations, protein partners, and proteolytic enzymes.
Unfortunately, the myriad of structural features, interaction surfaces, and post translational modifications on BRAF and its associated proteins make it challenging to understand how the kinase functions in normal physiology and how genetic mutations hijack the kinase and drive human cancer.
The Dana-Farber team generated purified forms of normal and mutant BRAF in addition to its key protein partners typically found in cells, and then assembled these to produce detailed molecular structures by cryo-electron microscopy. In addition, the researchers digested small aliquots of these complexes with a combination of proteolytic enzymes and then used the ZipChip on a Thermo Q-Exactive HF mass spectrometer to identify phosphorylation sites on BRAF and its co-regulatory protein partners. The rapid-fire, 10-minute injection-to injection throughput
provided by the ZipChip enabled the researchers to analyze hundreds of samples spanning different BRAF mutations, protein partners, and proteolytic enzymes. The multitude of ‘targeted’ and ‘unbiased’ CE-MS/ MS data sets enabled the researchers to read-out phosphorylation stoichiometry at key sites on BRAF which control its kinase activity and interaction with other co-regulatory proteins.
The Team combined their proteomic and structural data to better understand how BRAF mutations disrupt its normal cellular function. These results provide important insight to guide development of new, precision drugs which may improve clinical outcomes for patients with BRAF driven cancers.
To view the full article published in Nature Magazine click here.
[post_title] => Architecture of autoinhibited and active BRAF–MEK1–14-3-3 complexes
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H I G H L I G H T S
- CE-MS analysis for in-depth comparison of biosimilars
- The ZipChip® platform couples microfluidic capillary zone electrophoresis (CE) separation of intact proteins with direct electrospray ionization mass spectrometry (ESI-MS) analysis, facilitating the evaluation of drug product heterogeneity and comparison between originator and biosimilars.
- The high number of components confidently identified and quantified allows fast assessment of biologics similarity.
The ZipChip platform facilitates the evaluation of drug product heterogeneity and comparison between originator and biosimilars.
Materials & Methods
Sample Prep:
100 µg of each monoclonal antibody were buffer-exchanged according to the ZipChip protocol for intact antibody charge variant analysis with 0.5 mL spin-filters with 10KDa MWCO and concentrated to 0.5 mg/mL.
ZipChip Protocol Boosting Sensitivity for Intact Antibody Charge Variant Analysis Assay Kit Native Antibodies Kit Chip Type ZipChip HRN Field Strength 500 V/cm Run Time 15 minutes Mass Spec Type Thermo Scientific QExactive™ Plus Hybrid Quadrupole Orbitrap mass spectrometer
Introduction
According to the FDA “a biosimilar is a biological product that is highly similar to and has no clinically meaningful differences from an existing FDA approved reference product” and similarity is assessed “by extensively analyzing (i.e., characterizing) the structure and function of both the reference product and the proposed biosimilar” and only “minor differences between the reference product and the proposed biosimilar product in clinically inactive components are acceptable”1. Assessment of biosimilarity is usually performed by extensively analyzing the structure and function of both the reference product and the proposed biosimilar. Based on their high similarity with already approved drugs and their ability to lower the cost of current treatment, a shorter pathway has been designed for biosimilar approval under the Biologics Price Competition and Innovation (BPCI) Act, approved in the US in 2010. As a consequence, biopharmaceutical companies have interest in analytical tools that facilitate in-depth characterization of therapeutics to assess biosimilarity, in particular, the micro-heterogeneities present in the originator product. For this purpose, usually multiple orthogonal analyses are needed. For biosimilar approval the major effort is needed in terms of analytical rather than clinical studies (Fig 1).
In the present case study we evaluated the suitability of ZipChip platform to investigate biosimilarity by analyzing two commercially available infliximab drug products: Remicade the originator product, approved in the US in 1998; targeting TNF-α, it is indicated for the treatment of several autoimmune disease, including Crohn’s disease and its biosimilar, Inflectra, the first biosimilar approved by EU regulatory agencies in 2013.
Results
In the assessment of biosimilarity there is a need for bioanalytical tools that enable fast and confident evaluation of a candidate’s micro heterogeneity with increased dynamic range to verify that no critical quality attributes demonstrate significant differences from the originator.
Charge variant analysis is a regulatory requirement and is usually performed through CE or LC methodology to assess the charge variant profile2. Hyphenation of charge sensitive
Figure 1: Difference between approval process for originator and biosimilar therapeutic proteins.
separation methods with mass spectrometry has been shown to be a powerful characterization technique to understand the distribution and contribution of posttranslational modifications to the overall charge variant profile3,4. Infliximab drug products have a complex charge variant pattern, derived from the C-terminal lysine heterogeneity, with proteoforms presenting both, one or no terminal lysine residues5. In this complex pattern, low abundant proteoforms arising from N-glycan heterogeneity6, could easily be hidden below main species if only optical detection were used; MS hyphenation ensures low abundant species are not missed, especially with the high sensitivity attainable on the ZipChip platform.
Moreover, the minimal sample preparation required ensures no artificially induced modifications are introduced during the analysis, allowing a more reflective comparison of drug originator product and biosimilar candidates.
Figure 2 shows the electropherogram obtained from the Remicade drug product, with 3 main peaks identified and several minor species, all presenting complex glycoform heterogeneity, yielding a total of almost 100 proteoforms for both molecules, confidently identified with mass accuracies < 20 ppm. On the most acidic main peak (A), it is possible to observe a complex heterogeneity arising from deamidation and sialylation (peaks A1-A6). The same pattern is visible for peaks B and C even though most of the low abundant species migration times overlap with more abundant species in peak A and B, respectively. It is clear that in this case the evaluation of data acquired through optical detection only would not allow correct evaluation of all charge variants.
From the data obtained it was possible to generate evaluation of overall deamidation abundance, N-glycan profiling and lysine variants differences in the two monoclonal antibodies, including species as low as 0.1 %. In particular, Inflectra drug product is characterized by a higher level of C-terminal lysine cleavage and higher levels for terminal galactose on Fc N-glycan.
The excellent data quality and sensitivity, together with the short analysis time required by this platform, are key for a quick and in-depth screening and evaluation of biosimilarity across biologics.
Figure 2: Electropherogram obtained from CE-MS analysis on infliximab Remicade® drug product.
Authors:
Sara Carillo1, Craig Jakes, Florian Füssl, Jonathan Bones1,2
1. National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland. 2. School of Chemical and Bioprocess Engineering, University College Dublin, Dublin 4.
References
1. https://www.fda.gov/drugs/biosimilars/biosimilar-and-interchangeable-products 2. Sosic Z, Houde D Blum A, Carlage T, Lyubarskaya Y, Electrophoresis
2008; 29(21): 4368-4376. 3. Redman EA, Mellors JS, Starkey JA, Ramsey JM.
Anal Chem. 2016; 88(4): 2220-2226. 4. Füssl F, Cook K, Scheffler K, Farrell A,
Mittermayr S, Bones J. Anal Chem. 2018; 90(7): 4669-4676. 5. Redman EA, Batz
NG, Mellors JS, Ramsey JM. Anal Chem. 2015, 87(4): 2264-2272. 6. Marti SM,
Delporte C, Farrell A, Iglesias NN, McLoughlin N, Bones J. Analyst 2015, 140:
1442-1447.
[post_title] => Analysis to unravel biosimilarity of therapeutic proteins
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H I G H L I G H T S
- The demonstrated ZipChip® method is fast, simple, sensitive and uses the fully-automated microchip electrophoresis platform.
- Cell culture media can be directly analyzed using ZipChip after simple dilution with no further sample manipulation required.
- Amino acids can be monitored in less than 3 minutes using ZipChip, which is a faster alternative than conventional methods.
- Demonstrated ease of use for screening and monitoring amino acids in cell culture media.
Amino acids can be monitored in less than 3 minutes using ZipChip, which is a faster alternative than conventional methods.
Materials & Methods
Sample Prep:
Spent media samples were prepared following the ZipChip protocol for quantifying amino acids in growth media. Briefly, spent media was diluted 20x with LC-MS grade water and then 10x with sample diluent from the ZipChip Metabolites Kit.
Cell Line and Cell Culture Conditions:
CHO cells were adapted to grow in suspension. The culture experiments were initiated by seeding 0.3 x 106 cells/mL in 250 mL polycarbonate Erlenmeyer flasks containing 100 mL of chemically defined CHO cell culture media supplemented with 4mM of L-glutamine on day 0 in triplicate. All cultures were incubated at 37°C during the 10 days. Sampling was made daily by removing 1 mL aliquots, which were centrifuged, filtered and the supernatant stored at -20 °C for further analysis.
ZipChip Protocol Quantification of Amino Acids in Growth Media Assay Kit Metabolites Kit Chip Type ZipChip HS Field Strength 1000 V/cm Run Time 3 minutes Mass Spec Type Thermo Scientific QExactive™ Plus Hybrid Quadrupole Orbitrap mass spectrometer
Introduction
The biopharmaceutical industry has grown significantly over recent years with further expansion predicted based on packed pipelines of recombinant proteins, monoclonal antibodies and peptide drugs. Chinese Hamster Ovary (CHO) cells have been widely utilized for expression of recombinant proteins, which require media to support growth and therapeutic protein production1,2. The composition of cell culture media requires a blend of amino acids, vitamins, nucleosides, lipids, carbohydrates, trace elements and other components. Among these classes of compounds, amino acids are crucial as they are the constituent of proteins and are intermediates of many cellular metabolic pathways. As cells grow and produce therapeutic proteins certain amino acids are rapidly consumed. Monitoring the dynamic depletion of amino acids from culture media is important for media design and feeding strategy development. Supplementation with optimal amount of depleted amino acids is required to assure healthy growth, high productivity and to avoid formation of toxic or undesired metabolites3,4.
Various methods have been employed to analyze amino acids in spent media including LC-UV, LC-MS, GC-MS and CE-MS. However, these methods require sample preparation such as derivatization, solid phase extraction or microextraction or other steps in the experimental procedure prior to analysis. Derivatisation of some media components is required to enable optical detection using absorbance or fluorescence or for matrix simplification5, 6.
Miniaturized devices and microfluidic techniques offer the advantage of reduced analysis times and reduced consumption of reagents and therefore an environmentally friendly alternative to conventional methods.
In this study, we describe a simple, rapid, and automated microfluidic ZipChip ESI-MS method to monitor amino acids in spent media from an IgG1 producing CHO cell line using batch culture over a period of 10 days. The analysis facilitated identification of amino acids consumed during cell growth.
Results
During cell growth, amino acids are depleted from the culture media. Monitoring amino acid behaviour is crucial to understand the dynamic conditions and to adjust the concentration for each amino acid in the cell culture media to obtain the maximum possible production of monoclonal antibody. A simple, rapid and automated microfluidic
ZipChip ESI-MS method was developed to monitor amino acids in spent media from IgG1 producing CHO cells growing over a period of 10 days using batch culture.
The base peak electropherogram for the separation of 17 amino acids in water (reference standard) and in cell culture media employing ZipChip ESI-MS is shown in Figure 1. The separation is based on the difference of electrophoretic mobility, related to the charge and size of the analytes. The amino acid migration order observed was the highest positively charged amino acids migrate first, followed by the neutral amino acids and finally the acidic amino acids. The separation of 17 amino acids in 3 minutes showed symmetric peaks and good resolution. Some of the amino acids are unresolved, co-migration of certain amino acid pairs are commonly described in CE applications. However, with high resolution MS detection, co-migrating pairs can be easily distinguished based on differences in m/z.
The method was applied to study samples of conditioned cell culture media collected from flasks over the course of the 10-day batch culture. The culture media is rich in essential amino acids. Figure 2 shows the decline of signal intensity for the majority of the amino acids from day 0 to day 10.
All the amino acids can be quantified by this method. Figure 3 shows the patterns of leucine across the ten days of cell culture. As expected, the amount of the leucine is reduced during the 10-day culture.
Results
ZipChip ESI-MS facilitates rapid quantitative analysis of amino acid in cell culture media using an automated platform generating excellent separation within 3 minutes. The method provides good resolution, selectivity and requires minimal sample preparation and sample handling.
The ZipChip is suitable for screening and monitoring changes in cell culture media during therapeutic protein production with a short analysis time with reliable peak identification due to the coupling of CE with MS.
Figure 1: A mix of amino acids was diluted in CE buffer to optimize the separation via chip CE-ESI-MS. (a) Base peak electropherogram for a 500µM of amino acids mix standard.
Figure 2: Base peak electropherograms of cell culture growth from different days. (a) Day 1 and (b) Day 10
Figure 3: (a) Pattern of consumption of leucine during 10 days of cell culture growth (b) Extracted ion electropherograms of Leucine during 10 days of cell culture growth.
Authors:
Meire Ribeiro da Silva, Izabela Zaborowska, Sara Carillo, Jonathan Bones
National Institute for Bioprocessing Research & Training, Dublin, Ireland
References
1.https://www.fda.gov/drugs/biosimilars/biosimilar-and-interchangeable-products 2. Sosic Z, Houde D Blum A, Carlage T, Lyubarskaya Y, Electrophoresis
2008; 29(21): 4368-4376. 3. Redman EA, Mellors JS, Starkey JA, Ramsey JM.
Anal Chem. 2016; 88(4): 2220-2226. 4. Füssl F, Cook K, Scheffler K, Farrell A,
Mittermayr S, Bones J. Anal Chem. 2018; 90(7): 4669-4676. 5. Redman EA, Batz
NG, Mellors JS, Ramsey JM. Anal Chem. 2015, 87(4): 2264-2272. 6. Marti SM,
Delporte C, Farrell A, Iglesias NN, McLoughlin N, Bones J. Analyst 2015, 140:
1442-1447.
[post_title] => Automated monitoring of amino acids in cell culture media
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H I G H L I G H T S
- Performing highly selective charge variant separation of complex pharmaceuticals using the ZipChip® platform for native intact antibodies.
- Generation of excellent quality MS data which allows for in-depth analysis of also extremely low abundant charge variants.
- Demonstration of outstanding method sensitivity based on the identification of more than 200 Cetuximab isoforms with a sample consumption of only 2 ng of protein.
ZipChip is particularly beneficial in cases where only very limited quantities of sample are available.
Materials & Methods
Cetuximab was buffer exchanged to intact antibody sample diluent using Amicon Ultra-0.5 ML spin filters with a 30 kDa size cut-off to yield a 1 mg/mL concentration.
ZipChip Protocol Boosting Sensitivity for Intact Antibody Charge Variant Analysis Assay Kit ZipChip Native Antibody Kit Chip Type ZipChip HRN Field Strength 500 V/cm Run Time 20 minutes Mass Spec Type Thermo Scientific QExactive™ Plus Hybrid Quadrupole Orbitrap with Biopharma Option
Introduction
With the rise of precision medicine, the monoclonal antibody (mAb) market has expanded enormously during recent years. Monoclonal antibodies are large biomolecules that are produced by recombinant means and therefore can be highly heterogeneous. For therapeutic use, knowledge of this microheterogeneity is mandated by authorities to guarantee patient safety.
Charge variant analysis on the intact protein level represents an extremely powerful tool to distinguish protein isoforms based on differential surface charge, requiring minimal sample preparation and short analysis times. Recently, efforts were undertaken to combine charge variant separation of proteins with powerful detection modes such as mass spectrometry, creating new and exciting methods for protein characterisation1,2. Here, we explore the potential of ZipChip for the in-depth analysis of Cetuximab, a mAb that exceeds conventional mAbs in terms of complexity, as it contains four N-glycans, two in the conserved Fc region and another two in the Fab region (Figure 1) which can be complex and heavily sialylated. These additional Fab glycans are the cause of extremely high glycan heterogeneity and along with other charge altering modifications of the primary sequence are responsible for the complex peak pattern usually observed during charge variant analysis of Cetuximab. This charge variant and glycoform complexity makes Cetuximab a highly challenging molecule for in-depth characterization on the intact protein level.
Charge variant analysis is a regulatory requirement and is usually performed through CE or LC methodology to assess the charge variant profile2. Hyphenation of charge sensitive
Figure 1: General structure of Cetuximab. The Fc region contains simple, bi-antennary glycans also found on other mAbs whereas the Fab glycans can be larger, more complex and sialylated.
Results
Analysis of Cetuximab was performed using generic instrument parameters without the need for molecule specific optimization. The complex charge variant separation obtained can be seen in Figure 2. In total 8 Cetuximab charge variant peaks were distinguished, varying in abundance. Notably, all peaks are base-line separated which is important for subsequent MS-based isoform annotation and quantification. The averaged mass spectrum of the main charge variant peak is shown in blue in Figure 2. The spectral quality is excellent and allows for deconvolution and the characterization of even low abundant glycoforms. The averaged mass spectrum of the lowest abundant charge variant peak is shown in red in Figure 2. Spectral quality is sufficient for analysis, even for isoforms with relative abundance values of <1%.
Spectral deconvolution and analysis confirmed the expected charge variant migration in order of increasing negative protein net charge. The net charges are shown above each peak in Figure 3. Positive charges are caused by miss-cleaved C-terminal lysine, either on one or both heavy chains whereas negative charges are due to the presence
Figure 2: Charge variant separation of Cetuximab. The averaged spectra of the most (blue) and least (red) abundant charge variant peaks is shown
n the right panel.
of N-glycolyl neuraminic acid (NGNA) residues on the Fab glycans, as indicated in Figure 3. Net charges are ranging from +2 which corresponds to Cetuximab with 2 C-terminal lysine residues and no NGNA to -5 which represents Cetuximab without C-terminal lysine residues and five NGNA units attached. Importantly, each charge variant peak contains the characteristic glycosylation pattern of Cetuximab, giving rise to a range of isoforms found across the electropherogram.
More than 200 different isoforms were detected and quantified with relative abundance values of down to 0.1%. Considering that the run time required is only 15 minutes and a sample consumption of 2 ng, these results clearly demonstrate the power of the ZipChip for in-depth analysis of biopharmaceuticals. While ZipChip based CE-MS can be effectively employed for mAb characterization, it is in particular beneficial in cases where only very limited quantities of sample are available. This makes microfluidic CE-MS highly attractive for application in early stages of the biopharmaceutical production pipeline.
Figure 3: Assignment of protein net charge states to individual charge variant peaks. C-terminal lysine residues are labelled “K” and are shown in blue, N-glycolylneuraminic acids are labelled “NGNA” and are shown in turquois.
Authors:
Florian Füssl, Sara Carillo, Craig Jakes†, Jonathan Bones†
*National Institute for Bioprocessing Research and Training, Fosters Avenue,
Mount Merrion, Blackrock, Co. Dublin, Ireland.
† School of Chemical and Bioprocess Engineering, University College Dublin,
Dublin 4.
References
1. Redman, E. A.; Batz, N. G.; Mellors, J. S.; Ramsey, J. M. Anal Chem 2015, 87,
2264-2272. 2. Fussl, F.; Cook, K.; Scheffler, K.; Farrell, A.; Mittermayr, S.; Bones,
J. Anal Chem 2018, 90, 4669-4676. 3. 908devices. https://908devices.zendesk.
com, 2019.
[post_title] => Highly sensitive charge variant analysis of complex biopharmaceuticals
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H I G H L I G H T S
- Charge variant analysis of top selling monoclonal antibodies.
- Demonstrated ease of use, speed and depth of analysis obtained using the ZipChip® system, which couples microfluidic capillary zone electrophoresis (CE) separation of intact charge variants with direct electrospray ionization mass spectrometry (ESI-MS) analysis.
- The benefits of the ZipChip system include:
-Minimal sample preparation involved for many monoclonal antibody formulations.
-In the same fast analysis, a charge variant profile is obtained and is coupled with on-line MS identification in native conditions.
“Using the ZipChip system provides us with the ability to obtain charge variant separation in less than 15 minutes. This is a real advantage for our lab.„
- Sara Carillo
APPLICATION DEVELOPMENT TEAM LEADER, NIBRT
Materials & Methods
100 μg of each monoclonal antibody were prepared at a concentration of 0.5 mg/mL following the ZipChip protocol for intact antibody charge variant analysis.
ZipChip Protocol Boosting Sensitivity for Intact Antibody Charge Variant Analysis Assay Kit ZipChip Native Antibody Kit Chip Type ZipChip HRN Field Strength 500 V/cm Run Time 15 minutes Mass Spec Type Thermo Scientific QExactive™ Plus Hybrid Quadrupole Orbitrap with Biopharma Option
Introduction
The biopharmaceutical industry continues to develop mAb-based biotherapeutics for several applications and disease treatments. The complexity of these molecules creates a considerable challenge for the analytical technologies used to monitor the product quality attributes (PQAs) that need to be measured and controlled to guarantee safety and efficacy1. According to ICH guidelines, one of the features that needs to be monitored during biopharmaceutical development, and during approval and batch release post authorization, is the charge variant profile of a biotherapeutic. Standardized methods include the use of capillary electrophoresis using either zone or isoelectric focusing separation modes or the use of ion exchange liquid chromatography with UV detection (IEX-UV)2. The recent development of native mass spectrometry has facilitated the use of such separation techniques with Orbitrap mass spectrometry detection to obtain on-line MS identification of the charge variant proteoforms3,4. Native MS offers several advantages that include fast analysis and minimal sample preparation when combined with an upfront analytical separation, thus avoiding artificially-induced modification on the analyte.
ZipChip is a fast and reproducible tool for qualitative and quantitative charge variant analysis.
We tested the performance of the ZipChip system with a QExactive™ Plus Hybrid Quadrupole Orbitrap mass spectrometer to perform native MS experiments on the intact charge variants separated by ZipChip. The drug products tested in this work are rituximab, bevacizumab and trastuzumab.
Craig Jakes, PhD student, prepares the ZipChip system to begin a charge variant separation run
Results
The deep structural characterization of biotherapeutics is widely reported as a key step in new drug development and quality control to assess product safety and potency. Charge variant analysis is one of the product quality attributes that needs to be monitored during development and lot release to assess process robustness. In this study the ZipChip microfluidic capillary electrophoresis platform was applied for charge variant profiling of three top-selling drug products: rituximab, bevacizumab and trastuzumab.
For all the samples charge variant separation was obtained with little sample preparation and using a unique method for both CE separation and MS data acquisition. Moreover, the separation was obtained in less than 15 minutes in all the cases, providing a fast and reproducible tool for qualitative and quantitative charge variant analysis.
Figure 1: Zoom of the electropherograms obtained after ZipChip CE-ESI-MS analysis on bevacizumab (A), trastuzumab (B) and rituximab (C) drug products
Figure 1 shows the electropherograms obtained from the three drug products. For each molecule at least 10 proteoforms were identified, all presenting glycoform heterogeneity, with a total of up to 52 proteoforms identified for trastuzumab drug product. The use of QExactive™ plus mass spectrometer enables the fast acquisition of MS data using resolution setting of 35,000 at 200 m/z, routinely providing experimental masses with mass accuracies mostly below 20 ppm.
Due to the separation of the charge variants during the ZipChip separation, in addition to information on the glycoforms and C terminal lysine clipping, it was possible to determine pyro-glutamate formation and the presence of succinimide containing variants as well as deamidation
levels. These post-translational modifications are usually difficult to monitor due to the molecular mass of these variants being near isobaric to the main variant.
Overall, modifications with less than 0.1% abundancy have been confidently identified concluding that the ZipChip system is a powerful tool for biotherapeutics analysis.
Authors:
Sara Carillo, Craig Jakes, Jonathan Bones
National Institute for Bioprocessing Research & Training, Dublin, Ireland
References
1.Tassi M, De Vos J, Chatterjee S, Sobott F, Bones J, Eeltink S., J Sep Sci.,
2018; 41(1): 125-144. 2. Sosic Z, Houde D Blum A, Carlage T, Lyubarskaya Y,
Electrophoresis 2008; 29(21): 4368-4376. 3. Redman EA, Mellors JS, Starkey
JA, Ramsey JM. Anal Chem. 2016; 88(4): 2220-2226.
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Getting life-changing biologics to patients ASAP
HJB’s goal is to overcome scientific and technical hurdles that inhibit patients’ access to life-changing biologics. To do that, they aim to provide high quality, holistic, and reliable services to innovative biopharma companies.
So how exactly do they hit that lofty goal? The contract development and manufacturing organization (CDMO) based in Zhejiang, China, employs cutting-edge technology that substantially accelerates R&D processes, reduces costs, and fast-tracks commercialization. They offer biopharma clients access to complete, flexible, and reliable development and manufacturing services that include cell line development, bioprocess development, analytical method development, formulation development and GMP manufacturing for all clinical phases and commercial production.
Overcoming hurdles in cell culture media analysis
CHALLENGE: The waiting game
Prior to HJB’s integration of the REBEL™ cell culture media analyzer into their processes, the upstream process development team, led by Vice President Lisa Zheng, sent media samples off to HJB’s analytical sciences group for amino acid analysis. But relying on another team for results meant they were waiting to make decisions and take next steps in their media development processes.
In their search to accelerate R&D and reduce costs, HJB discovered the power of REBEL and ZipChip.
SOLUTION: Answers at-line, anytime
Lisa’s team purchased a REBEL and set it up right next to their bioreactors. They found the device space-friendly and easy to install. Consumables were easy to maintain, and operation was so simple that even their non-technical analysts could conduct analysis. Samples were measured quickly, and with results in less than 10 minutes, scientists were making decisions in near-real time. REBEL is now used routinely for quick analysis to support accelerated media and process improvements.
CONCLUSION: A new milestone in productivity
Because REBEL lets the upstream process development group analyze many samples in a short period of time, the group accumulates a large amount of amino acid data just through daily testing. This lets them quickly investigate deviations during product development, carry out process development/optimization based on REBEL data, and better understand the impacts on culture performance and Product Quality Attributes (PQAs) from process parameter changes.
REBEL was also key in helping HJB accelerate media and process development, which is essential for the development of highly intensified, continuous cell culture perfusion. Earlier this year, the company announced that its continuous perfusion cell culture platform has achieved industry-leading volumetric productivity of greater than 6 g/L per day while maintaining process and product quality attributes in a state of control for a 4-week culture.
“REBEL can analyze a large number of samples in a short period of time to provide valuable data to accelerate cell culture process development. Rather than obtaining amino acid profiles after the run is completed, REBEL provides real-time data for us to make process decisions while the experiments are in progress to maximize successful outcomes.”
-Dr. Christopher Hwang, CTO of HJB
Overcoming hurdles in cell culture media analysis
CHALLENGE: A fair share of challenges with LC
Liming Shi is the VP in charge of the Analytical Science and Quality Control Departments, and responsible for Product & Process Development Operation. HJB’s Analytical Science Department develops and optimizes analytical methods, and provides analytical support for cell lines upstream, downstream, and formulation process development. The group applies the most advanced analytical techniques to perform large molecule characterization, including analysis of mAbs, ADCs, and bispecific antibodies.
Liming’s team was looking for difficult-to-separate charge variants, which is a challenge with LC-based separations. They were also bumping into UHPLC challenges, such as poor separation and analysis with limited sample volumes. Furthermore, their UHPLC-MS separation and analysis had a turn-around time that was too long.
SOLUTION: In comes ZipChip
In their search for analytical tools that could provide higher-quality data and get them to results faster, the Analytical Services team put ZipChip® straight to work.
Because ZipChip enables rapid measurement of Critical Quality Attributes (CQAs) of native and intact monoclonal antibodies (mAbs), the team used it to characterize intact mAbs and assess the root cause of increases in acidic and basic variants under stress at high temperature. The antibody was analyzed at multiple levels, including its intact state under native conditions and subunit and peptide levels. They used the same ZipChip system and method for both analyses, and were able to determine charge heterogeneity speciation and identification in under 10 minutes. This new workflow was 100 times faster with less prep compared to conventional LC, which required them to harvest fractions.
It was also easy to start integrating ZipChip into their other laboratory workflows as minimal method development was needed for different types of proteins. The team noted this was a big benefit when it came to analysis of native proteins.
Liming Shi -
VP of Analytical Science and Quality Control Departments, Product & Process Development Operation
“ZipChip is a capable platform for multiple CQA assays, and for charge variant analysis it is very straightforward to get separation and identification information with minimal method development needed for different types of proteins, especially for the analysis in native conditions. The sample preparation is simple and straightforward, and the sample requirements are at nanogram levels.”
CONCLUSION: ZipChip + MS goes mainstream
The Analytical Science and Quality Control groups at HJB now have ZipChip systems coupled to their Q Exactive and other Thermo Fisher Scientific mass specs. They’ve adopted ZipChip into their intact antibody analysis workflows (denatured and native conditions) and their subunit and peptide mapping analysis as it gives them very clear and in-depth information on antibody molecules. One of the stand-out features in Liming’s opinion is having an online platform that provides straightforward separation and identification of charge variants based on different mechanisms.
The Analytical Science team recently published their work using ZipChip in Analytical Biochemistry, Volume 625, 15 July 2021, 114214: Characterization and monitoring of charge variants of a recombinant monoclonal antibody using microfluidic capillary electrophoresis-mass spectrometry.
[post_title] => REBEL + ZipChip help HJB Overcome Scientific Hurdles and Optimize Processes in Biotherapeutic Development
[post_excerpt] => In their search to accelerate R&D and reduce costs, HJB discovered the power of REBEL and ZipChip.
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Case Studies
REBEL + ZipChip help HJB Overcome Scientific Hurdles and Optimize Processes in Biotherapeutic Development
In their search to accelerate R&D and reduce costs, HJB discovered the power of REBEL and ZipChip.
Architecture of autoinhibited and active BRAF–MEK1–14-3-3 complexes
Analysis to unravel biosimilarity of therapeutic proteins
Automated monitoring of amino acids in cell culture media
Highly sensitive charge variant analysis of complex biopharmaceuticals
In-depth analysis of top selling biotherapeutics
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Advancements in bioprocess characterization technologies have progressed rapidly, broadening the range of readily accessible process information. While dissolved gases, pH, and temperature have been measurable in- or on-line for some time, crucial nutrients and metabolites are typically monitored via manual sampling and at- or off-line analysis.
This poster was a collaboration with Terumo Blood and Cell Technologies and was part of the poster program at the 2024 ISCT EU Conference.
[post_title] => Enhancing Cell and Gene Therapy Manufacturing Processes by In-line, On-line and At-line Bioprocess Analytical Technologies
[post_excerpt] => The 908 Devices, on-, and in-line analytical solutions contribute to an enhanced understanding, characterization, and control of bioprocesses in development and manufacturing of cell and gene therapies.
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[post_title] => Microchip CE-MS Analysis of Nucleic Acids – From Characterization of Synthetic Oligonucleotides to Sequence Mapping of RNAs
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[post_title] => Investigation of Drug-To-Antibody Ratio for FORCE Oligonucleotide Conjugates Using Microchip CE-MS
[post_excerpt] => ZipChip microchip CE-MS resolves DAR species electrophoretically and enables generation of superior MS spectra. This poster was presented as a collaboration with Dyne Therapeutics at ASMS 2023 as poster WP045.
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[post_title] => In-depth Characterization of Adeno-Associated Viruses using Microchip Capillary Electrophoresis Coupled with Mass Spectrometry
[post_excerpt] => Rapid microchip CE-MS can be utilized for the in-depth characterization of AAVs including viral capsid protein analysis and peptide mapping. This poster was presented as a collaboration with NIBRT at ASMS 2023 as poster WP730.
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[post_title] => Rapid In-Depth Characterization of Biologics by Microchip CE-MS: mAbs, AAVs and Nucleic Acids
[post_excerpt] => Microchip capillary electrophoresis (CE) coupled with mass spectrometry (MS) has revolutionized the characterization of therapeutic modalities in biopharmaceuticals. Here, we present the applications of an integrated workflow that couples the microfluidic CE system (ZipChip) with MS in characterization of biotherapeutics. This was poster A03 at BPS Europe 2024.
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[post_title] => 5’ mRNA Analysis By Microchip CE-MS Using an Internal Cleavage Motif for RNase H Digestion
[post_excerpt] => The subject of this poster was the investigation of an internal DNA motif to produce a shorter digestion product while simultaneously maintaining the length of the probe at 25nt for hybridization specificity. The shorter oligo results in easier detection using Microchip CE-MS approach. This poster was presented at ASMS 2023 as poster THP575.
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[ping_status] => closed
[post_password] =>
[post_name] => 5-mrna-analysis-by-microchip-ce-ms
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-26 14:32:16
[post_modified_gmt] => 2024-03-26 18:32:16
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[guid] => https://908devices.com/?post_type=resource&p=23828
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[6] => WP_Post Object
(
[ID] => 23830
[post_author] => 15
[post_date] => 2024-03-25 10:15:07
[post_date_gmt] => 2024-03-25 14:15:07
[post_content] =>
[post_title] => Improved Performance of Phosphopeptides Characterization Using Online Capillary Electrophoresis Coupling with Ion Mobility Mass Spectrometry (CE-IM-MS)
[post_excerpt] => This poster will highlight a novel strategy for characterization of therapeutic peptide that was developed based on microchip CE-MS pure liquid separation and ion mobility gas phase separation, combing with mass spectrometry identification. The poster was presented in collaboration with Ningbo University at ASMS 2023 as poster THP609.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => improved-performance-of-phosphopeptides-characterization
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-25 10:15:12
[post_modified_gmt] => 2024-03-25 14:15:12
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[guid] => https://908devices.com/?post_type=resource&p=23830
[menu_order] => 114
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[7] => WP_Post Object
(
[ID] => 23820
[post_author] => 15
[post_date] => 2024-03-25 09:27:20
[post_date_gmt] => 2024-03-25 13:27:20
[post_content] =>
[post_title] => Rapid Identification of Conjugation Sites in Antibody Drug Conjugates Using Microchip Capillary Electrophoresis Coupled with Mass Spectrometry
[post_excerpt] => CE/MS method is optimized with NISTmAb digest as a reference control. Good MS/MS coverage of NISTmAb is obtained with narrow peak width and fast separation. This poster was presented as a collaboration with Bruker at ASMS 2023 as poster WP041.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => rapid-identification-of-conjugation-sites-in-antibody-drug
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-25 09:28:04
[post_modified_gmt] => 2024-03-25 13:28:04
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23820
[menu_order] => 116
[post_type] => resource
[post_mime_type] =>
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[8] => WP_Post Object
(
[ID] => 23818
[post_author] => 29
[post_date] => 2024-03-25 09:13:48
[post_date_gmt] => 2024-03-25 13:13:48
[post_content] =>
[post_title] => Conformation of Native Antibody-Drug Conjugate Charge Variants Revealed by Microchip Capillary Electrophoresis Coupled with Trapped Ion Mobility
[post_excerpt] => The combination of microchip capillary electrophoresis and trapped ion mobility enables a rapid, informative method for native monoclonal antibody and antibody-drug conjugate charge variant analysis. This poster was a collaboration with Bruker and NIH and presented at ASMS 2023 as poster WP040.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => conformation-of-native-antibody-drug-conjugate-charge-variants
[to_ping] =>
[pinged] =>
[post_modified] => 2024-05-07 14:29:50
[post_modified_gmt] => 2024-05-07 18:29:50
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23818
[menu_order] => 117
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[9] => WP_Post Object
(
[ID] => 23798
[post_author] => 15
[post_date] => 2024-03-21 11:02:11
[post_date_gmt] => 2024-03-21 15:02:11
[post_content] =>
[post_title] => Fast Screening and Characterization of Therapeutic Peptide by Online Capillary Electrophoresis - Mass Spectrometry (CE-MS)
[post_excerpt] => Presenting a novel strategy for characterization of therapeutic peptides was developed based on microchip CE-MS and linkage-specific derivatization. This poster was presented in collaboration with Ningbo University at ASMS 2023 as poster TP544.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => fast-screening-and-characterization-of-therapeutic-peptide
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 11:02:12
[post_modified_gmt] => 2024-03-21 15:02:12
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23798
[menu_order] => 118
[post_type] => resource
[post_mime_type] =>
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[filter] => raw
)
[10] => WP_Post Object
(
[ID] => 23796
[post_author] => 15
[post_date] => 2024-03-21 10:52:20
[post_date_gmt] => 2024-03-21 14:52:20
[post_content] =>
[post_title] => Continuing the Investigation of Microchip Capillary Electrophoresis Coupled with Mass Spectrometry in the Bottom-Up Characterization of Progressively Larger RNAs
[post_excerpt] => Exploring the merits of capillary-zone electrophoresis (CZE)-mass spectrometry to analyze digestion mixtures of large RNAs and attain sequence information. This poster was presented as a collaboration with UConn and Bruker at ASMS 2023 and was poster TP532.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => continuing-the-investigation-of-microchip-capillary-electrophoresis
[to_ping] =>
[pinged] =>
[post_modified] => 2024-08-01 14:18:20
[post_modified_gmt] => 2024-08-01 18:18:20
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23796
[menu_order] => 119
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[11] => WP_Post Object
(
[ID] => 23794
[post_author] => 15
[post_date] => 2024-03-21 10:38:52
[post_date_gmt] => 2024-03-21 14:38:52
[post_content] =>
[post_title] => Determination of ATP, ADP, AMP, and Adenosine Levels by Microchip Capillary Electrophoresis Coupled with High Resolution Mass Spectrometry
[post_excerpt] => In this study, we present a novel ZipChip coupled with a QEHF for adenosine nucleotide analysis. This poster was presented at ASMS 2023 as poster TP489.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => determination-of-atp-adp-amp-and-adenosine-levels
[to_ping] =>
[pinged] =>
[post_modified] => 2024-08-01 14:18:43
[post_modified_gmt] => 2024-08-01 18:18:43
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23794
[menu_order] => 120
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[12] => WP_Post Object
(
[ID] => 23787
[post_author] => 15
[post_date] => 2024-03-21 10:20:58
[post_date_gmt] => 2024-03-21 14:20:58
[post_content] =>
[post_title] => Sample to Result Workflow for the Investigation of Biosimilars vs Innovator Cetuximab by Charge Variant Analysis using Microchip CE-MS
[post_excerpt] => This poster was presented in collaboration with Protein Metrics and highlights how the ZipChip coupled to HRMS provides a streamlined analytical approach to rapidly assess the heterogeneity of the investigated therapeutic proteins through the comprehensive interrogation of the different charge variants and their intact MS spectra. This poster was presented at ASMS 2023 as poster TP 384.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => sample-to-result-workflow-for-the-investigation-of-biosimilars-vs-innovator-cetuximab
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 10:20:58
[post_modified_gmt] => 2024-03-21 14:20:58
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23787
[menu_order] => 122
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[13] => WP_Post Object
(
[ID] => 23785
[post_author] => 15
[post_date] => 2024-03-21 10:15:48
[post_date_gmt] => 2024-03-21 14:15:48
[post_content] =>
[post_title] => Peptide Mapping Workflow for Direct Microchip CE-MS Analysis of Biopharmaceuticals
[post_excerpt] => In this study we demonstrated the suitability of a modified digestion workflow for peptide mapping analysis via CE separation. This poster was presented at ASMS 2023 in collaboration with NIBRT. It was poster MP619.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => peptide-mapping-workflow-for-direct-microchip
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 10:15:49
[post_modified_gmt] => 2024-03-21 14:15:49
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23785
[menu_order] => 123
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[14] => WP_Post Object
(
[ID] => 23781
[post_author] => 15
[post_date] => 2024-03-21 09:57:40
[post_date_gmt] => 2024-03-21 13:57:40
[post_content] =>
[post_title] => Combining Capillary Electrophoresis and Trapped Ion Mobility Spectrometry Mass Spectrometry to Analyze Epitranscriptomic Marks Mediating Virus-Host Interactions in Infectious Diseases
[post_excerpt] => This poster evaluates the merits of capillary-zone electrophoresis (CZE)-MS. This poster was presented at ASMS 2023 as a collaboration with UCONN. It was poster MP548.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => combining-capillary-electrophoresis-and-trapped-ion-mobility-spectrometry-mass-spectrometry
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 09:57:41
[post_modified_gmt] => 2024-03-21 13:57:41
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23781
[menu_order] => 124
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[15] => WP_Post Object
(
[ID] => 23779
[post_author] => 15
[post_date] => 2024-03-21 09:38:33
[post_date_gmt] => 2024-03-21 13:38:33
[post_content] =>
[post_title] => Use of Microchip Capillary Electrophoresis – Mass Spectrometry for Automated Rapid Measurement of Enzyme Reaction Kinetics
[post_excerpt] => This approach allows rapid measurement of enzymatic cleavage dynamics in real time. It can be applied to almost any enzyme substrate reaction and is highly tolerable to various buffers, thanks to the CE Chip-MS configuration. This poster was presented at ASMS 2023 with collaborators as poster MP547.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => use-of-microchip-capillary-electrophoresis
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 09:55:53
[post_modified_gmt] => 2024-03-21 13:55:53
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23779
[menu_order] => 125
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[16] => WP_Post Object
(
[ID] => 23776
[post_author] => 15
[post_date] => 2024-03-21 09:29:35
[post_date_gmt] => 2024-03-21 13:29:35
[post_content] =>
[post_title] => Rapid CE-MS Analysis of Released N-Glycan: Optimized Workflow for Direct CE Compatibility
[post_excerpt] => N-glycan profiling is a critical step of biopharmaceuticals analysis as N-glycans play an important part in both activity and safety of monoclonal antibodies and other glycosylated biotherapeutics. This poster was presented at ASMS 2023 as poster MP291.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => rapid-ce-ms-analysis-of-released-n-glycan
[to_ping] =>
[pinged] =>
[post_modified] => 2024-03-21 09:35:32
[post_modified_gmt] => 2024-03-21 13:35:32
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23776
[menu_order] => 128
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[17] => WP_Post Object
(
[ID] => 25226
[post_author] => 29
[post_date] => 2024-07-09 11:04:45
[post_date_gmt] => 2024-07-09 15:04:45
[post_content] =>
Understanding of cell culture metabolism in biologics manufacturing has increased tremendously in the past decade; however, the lack of feasible methods of high coverage, quantitative information on the interplay between nutrients and metabolites has hindered the efforts to fully characterize processes in the different biological phases. Several published models support cell growth phase but lack the kinetic information to be applicable to the phases that follow. We discuss a method for measuring hundreds of metabolites in dozens of pathways that streamlines sample preparation from spent media and cells, enables fast data-analysis times and supports quantitative analysis of key metabolites to gain far richer and more reliable data sources for process characterization, modelling and understanding. The method uses microchip CE/MS and incorporates stable isotope internal standards for both quantitation and migration time indexing. This work explores the information that can be obtained from the extracellular growth media and intracellular metabolites extracted from the CHO cells.
The poster was on display at Bioprocessing Summit, and ESACT #253 in 2024.
[post_title] => In-depth Metabolic Characterization at Different Phases of Cell Culture with a Turnkey CE-ESI-HRMS Workflow
[post_excerpt] => Learn about a turnkey CE-MS metabolomics workflow solution used to characterize media metabolites and intracellular metabolites of CHO bioreactors. The poster was on display at Bioprocessing Summit, and ESACT #253 in 2024.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => correlation-of-soluble-metabolites-with-lactate-switch-observed-in-hek293-culture-using-novel-ce-ms-metabolomics-workflow
[to_ping] =>
[pinged] =>
[post_modified] => 2024-11-25 16:13:49
[post_modified_gmt] => 2024-11-25 21:13:49
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=25226
[menu_order] => 130
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[18] => WP_Post Object
(
[ID] => 26149
[post_author] => 15
[post_date] => 2024-10-11 13:20:46
[post_date_gmt] => 2024-10-11 17:20:46
[post_content] =>
Advancements in bioprocess characterization technologies have progressed rapidly, broadening the range of readily accessible process information. While dissolved gases, pH, and temperature have been measurable in- or on-line for some time, crucial nutrients and metabolites are typically monitored via manual sampling and at- or off-line analysis. This poster was developed with Terumo Blood and Cell Technologies and was showcased at BPI USA and Advanced Therapies 2024.
[post_title] => Innovations in Analytical Technology for Cell & Gene Therapy Manufacturing
[post_excerpt] => This poster highlights our at-, on-, and in-line analytical solutions that contribute to an enhanced understanding, characterization, and control of bioprocesses in the development and manufacturing of cell and gene therapies.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => innovations-in-analytical-technology-for-cell-gene-therapy-manufacturing
[to_ping] =>
[pinged] =>
[post_modified] => 2024-11-18 15:32:59
[post_modified_gmt] => 2024-11-18 20:32:59
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=26149
[menu_order] => 132
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[19] => WP_Post Object
(
[ID] => 25218
[post_author] => 29
[post_date] => 2024-07-09 10:54:09
[post_date_gmt] => 2024-07-09 14:54:09
[post_content] =>
The introduction of the ZipChip Intact Antibodies Reagent Kit marked a significant milestone in the ZipChip portfolio, enabling the first tailored solution of charge variant analysis (CVA) of monoclonal antibodies on the CE-MS platform. However, there was room for enhancement in preserving the native conformation of the proteins under analysis, as well as in enhancing the separation between variant peaks. The next generation kits, the Native Antibodies Kit and Charge Variant TOF Kit, significantly improved peak separation and producing spectra more indicative of native protein structures. This was poster # TP642 at ASMS 2024.
[post_title] => Optimized Native Conditions for Charge Variant Analysis Using a Microfluidic CE-MS Platform Yield Higher Quality Data and Enables Analysis of Lower Titer Samples
[post_excerpt] => Learn about the improvements made in the new Charge Variant Analysis kit, compared to the previous Intact Antibodies and Native Antibodies kits. ASMS 2024.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => optimized-native-conditions-for-charge-variant-analysis-using-a-microfluidic-ce-ms-platform-yield-higher-quality-data-and-enables-analysis-of-lower-titer-samples
[to_ping] =>
[pinged] =>
[post_modified] => 2024-09-23 12:16:29
[post_modified_gmt] => 2024-09-23 16:16:29
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=25218
[menu_order] => 133
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[20] => WP_Post Object
(
[ID] => 25213
[post_author] => 29
[post_date] => 2024-07-09 10:28:00
[post_date_gmt] => 2024-07-09 14:28:00
[post_content] =>
As the field of bioprocessing continues to mature, there is a need for better analytics applied closer to the bioprocess. We have seen recent advancements in the technology used to measure small molecules within bioreactors (glucose/lactate, amino acids), but there is also a need to directly measure the critical quality attributes of the proteins being produced. To that end, we are assessing the ability of microfluidic CE-ESI-MS to perform charge variant - MS analysis of proteins pulled directly from a bioreactor. We know that excessive salt concentrations can cause band-broadening for this separation, so the key question is whether a desalting step is required, or if a simple dilute and shoot approach can work, and how much information can be obtained. Here we compare a simple dilute and shoot method to a method that buffer exchanges the sample before ZipChip analysis and demonstrates the use of this method for analysis of 4 different bioreactor growth conditions. This poster was on display at ASMS 2024 as poster # TP638.
[post_title] => Charge Variant Analysis of IgGs Directly from Cell Culture Supernatant Using Microfluidic CE-ESI-MS
[post_excerpt] => This poster describes the application of Microfluidic CE-ESI-MS for analyzing protein charge variants directly from a bioreactor. It investigates the feasibility of a "dilute and shoot" approach compared to a buffer exchange method for sample preparation. ASMS 2024.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => charge-variant-analysis-of-iggs-directly-from-cell-culture-supernatant-using-microfluidic-ce-esi-ms
[to_ping] =>
[pinged] =>
[post_modified] => 2024-09-23 12:20:01
[post_modified_gmt] => 2024-09-23 16:20:01
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=25213
[menu_order] => 134
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[21] => WP_Post Object
(
[ID] => 25245
[post_author] => 15
[post_date] => 2024-07-09 14:32:05
[post_date_gmt] => 2024-07-09 18:32:05
[post_content] =>
The biopharmaceutical industry is increasingly motivated to improve process control, understanding, and repeatability using process analytical technologies (PAT). More specifically, Raman-based optical, in-line measurement methods have shown promise as a way to monitor critical performance parameters, including glucose and lactate concentrations, in real-time. Recently, this has translated to the implementation of Raman-based glucose feeding strategies to modulate productivity and product quality in Chinese hamster ovary (CHO) cell cultures. The current work implements a novel Raman-based solution that does not require any chemometric models for glucose, lactate and biomass measurements. MAVERICK was directly, without any training in the specific bioprocess, deployed to dynamically control lactate feeding. We compared feeding strategies as means of modulating product quality and improving culture conditions, primarily through decreased ammonia accumulation. This was poster #358 at ESACT 2024 and was also highlighted at the Bioprocessing Summit 2024.
[post_title] => Raman-based Lactate Feeding Impact on Culture Characteristics and Product Quality in Mammalian Cell Cultures
[post_excerpt] => This research explores Raman spectroscopy for real-time glucose monitoring and feeding strategies in CHO cell cultures, aiming to improve product quality.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => raman-based-lactate-feeding-impact-on-culture-characteristics-and-product-quality-in-mammalian-cell-cultures
[to_ping] =>
[pinged] =>
[post_modified] => 2024-08-23 14:16:28
[post_modified_gmt] => 2024-08-23 18:16:28
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=25245
[menu_order] => 142
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[22] => WP_Post Object
(
[ID] => 23438
[post_author] => 15
[post_date] => 2024-02-20 17:24:46
[post_date_gmt] => 2024-02-20 22:24:46
[post_content] =>
[post_title] => Improving Cell and Gene Therapy Manufacturing Processes by Automated On-line and In-line Bioprocess Analytical Technologies
[post_excerpt] => Recent advances in cell and gene therapy bioprocess analytical technology regimes, approaches to de novo analytical modeling/calibration of these systems, and how their informing power can be leveraged with the increasing sophistication of bioprocess metabolic models for predictive bioprocess optimization. View our poster from the 2024 Advanced Manufacturing Cell and Gene Therapies Conference. This was poster 29. This poster was also on display as #843 at ISCT 2024.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => improving-cell-gene-therapy-manufacturing
[to_ping] =>
[pinged] =>
[post_modified] => 2024-07-08 13:01:02
[post_modified_gmt] => 2024-07-08 17:01:02
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=23438
[menu_order] => 146
[post_type] => resource
[post_mime_type] =>
[comment_count] => 0
[filter] => raw
)
[23] => WP_Post Object
(
[ID] => 6117
[post_author] => 20
[post_date] => 2022-06-11 02:59:33
[post_date_gmt] => 2022-06-11 06:59:33
[post_content] =>
[post_title] => Bottom-Up Analysis of Progressively Larger RNAs by CZE-MS and CZE-MS/MS
[post_excerpt] => In this poster from ASMS 2022, researchers from University of Connecticut and Bruker showed that capillary zone electrophoresis performed on a chip proved to be an effective alternative to liquid chromatography as a front-end separation system to analyze both pure samples and complex digestion mixtures of nucleic acids.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => zipchip-poster-asms2022-uconn
[to_ping] =>
[pinged] =>
[post_modified] => 2023-12-26 12:43:57
[post_modified_gmt] => 2023-12-26 17:43:57
[post_content_filtered] =>
[post_parent] => 0
[guid] => https://908devices.com/?post_type=resource&p=6117
[menu_order] => 192
[post_type] => resource
[post_mime_type] =>
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[24] => WP_Post Object
(
[ID] => 6115
[post_author] => 20
[post_date] => 2022-06-11 02:52:58
[post_date_gmt] => 2022-06-11 06:52:58
[post_content] =>
[post_title] => Multi-level Expeditious Characterization of Adeno-associated Virus (AAV) Capsid Proteins Using ZipChip Microfluidic CE Coupled with HRMS
[post_excerpt] => In this poster from ASMS 2022, scientists from Thermo Fisher Scientific and 908 Devices present a novel approach using microchip CE-MS for the characterization of AAVs.
[post_status] => publish
[comment_status] => closed
[ping_status] => closed
[post_password] =>
[post_name] => zipchip-poster-asms2022-thermo
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[post_title] => A Sensitive and High-Throughput Assay for Analyzing Peptide Exchanged MHCl by High-Resolution Native CZE-MS
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[post_title] => Combining μSPE ZipChip CE with PRM-LIVE on a timsTOF Pro for Warp-speed Selectivity Profiling of Deubiquitinase (DUB) Small Molecule Inhibitors
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[post_title] => Microchip CE-MS Analysis of Sialylated N-glycan Linkage Isomers
[post_excerpt] => In this ASMS 2022 poster, Fudan University researchers show a ZipChip-based strategy for qualitative and quantitative analysis of sialylated N-glycan linkage isomers was developed.
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[post_title] => CE-MS of Oligonucleotides using a ZipChip
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[post_title] => Rapid Characterization of Oligos and Related Impurities Using Microchip CE-MS without Ion-pairing Reagents
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[post_title] => Glycosylation Profiling of mAB Subunits with CZE-MS
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[post_title] => Pushing the Boundaries of Speed and Sensitivity in Proteomics
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[post_title] => A Broad Application of Migration Time Indexing and Metabolite Libraries to Evaluate Quantitative Coverage in Microchip CE-MS Metabolomics
[post_excerpt] => In the field of metabolomics, microchip capillary electrophoresis (mCE) has a number of potential advantages over traditional liquid chromatography (LC), including higher throughput due to decreased separation times, increased separation efficiency, and lower required sample volumes.
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[post_title] => A Microchip CE-MS Based End-to-End Solution for the Analysis of Synthetic Oligonucleotides
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[post_title] => Microchip CE-MS with Integrated Solid Phase Extraction
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[post_title] => Microchip CE-MS of Oligonucleotides: What Can It Separate?
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[post_title] => Fast Chip-CEMS for Characterization of Protein/PTM Changes
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[post_title] => Combined Top-down and Bottom-up Proteomics
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[post_title] => ZipChip for Glycomics and Glycoproteomics
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[post_title] => Fast and Efficient Biopharmaceutical Characterization
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[post_title] => Characterization of charge heterogeneity of mAb
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[post_title] => Rapid, Multi-level Analysis of Complex Proteins
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[post_title] => Oligonucleotide Analysis via Microfluidic CE-MS
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[post_title] => Advancing mAb Characterization with Microchip CE-MS Couples to a PASEF Enabled QTOF
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Advancements in bioprocess characterization technologies have progressed rapidly, broadening the range of readily accessible process information. While dissolved gases, pH, and temperature have been measurable in- or on-line for some time, crucial nutrients and metabolites are typically monitored via manual sampling and at- or off-line analysis.
This poster was a collaboration with Terumo Blood and Cell Technologies and was part of the poster program at the 2024 ISCT EU Conference.
[post_title] => Enhancing Cell and Gene Therapy Manufacturing Processes by In-line, On-line and At-line Bioprocess Analytical Technologies
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Posters
Enhancing Cell and Gene Therapy Manufacturing Processes by In-line, On-line and At-line Bioprocess Analytical Technologies
The 908 Devices, on-, and in-line analytical solutions contribute to an enhanced understanding, characterization, and control of bioprocesses in development and manufacturing of cell and gene therapies.
Microchip CE-MS Analysis of Nucleic Acids – From Characterization of Synthetic Oligonucleotides to Sequence Mapping of RNAs
This work will showcase how the ZipChip enables a simple and rapid workflow for the analysis of nucleic acids with noion-pairing reagents needed This poster was highlighted at the 2024 TIDES Conference.
Investigation of Drug-To-Antibody Ratio for FORCE Oligonucleotide Conjugates Using Microchip CE-MS
ZipChip microchip CE-MS resolves DAR species electrophoretically and enables generation of superior MS spectra. This poster was presented as a collaboration with Dyne Therapeutics at ASMS 2023 as poster WP045.
In-depth Characterization of Adeno-Associated Viruses using Microchip Capillary Electrophoresis Coupled with Mass Spectrometry
Rapid microchip CE-MS can be utilized for the in-depth characterization of AAVs including viral capsid protein analysis and peptide mapping. This poster was presented as a collaboration with NIBRT at ASMS 2023 as poster WP730.
Rapid In-Depth Characterization of Biologics by Microchip CE-MS: mAbs, AAVs and Nucleic Acids
Microchip capillary electrophoresis (CE) coupled with mass spectrometry (MS) has revolutionized the characterization of therapeutic modalities in biopharmaceuticals. Here, we present the applications of an integrated workflow that couples the microfluidic CE system (ZipChip) with MS in characterization of biotherapeutics. This was poster A03 at BPS Europe 2024.
5’ mRNA Analysis By Microchip CE-MS Using an Internal Cleavage Motif for RNase H Digestion
The subject of this poster was the investigation of an internal DNA motif to produce a shorter digestion product while simultaneously maintaining the length of the probe at 25nt for hybridization specificity. The shorter oligo results in easier detection using Microchip CE-MS approach. This poster was presented at ASMS 2023 as poster THP575.
Improved Performance of Phosphopeptides Characterization Using Online Capillary Electrophoresis Coupling with Ion Mobility Mass Spectrometry (CE-IM-MS)
This poster will highlight a novel strategy for characterization of therapeutic peptide that was developed based on microchip CE-MS pure liquid separation and ion mobility gas phase separation, combing with mass spectrometry identification. The poster was presented in collaboration with Ningbo University at ASMS 2023 as poster THP609.
Rapid Identification of Conjugation Sites in Antibody Drug Conjugates Using Microchip Capillary Electrophoresis Coupled with Mass Spectrometry
CE/MS method is optimized with NISTmAb digest as a reference control. Good MS/MS coverage of NISTmAb is obtained with narrow peak width and fast separation. This poster was presented as a collaboration with Bruker at ASMS 2023 as poster WP041.
Conformation of Native Antibody-Drug Conjugate Charge Variants Revealed by Microchip Capillary Electrophoresis Coupled with Trapped Ion Mobility
The combination of microchip capillary electrophoresis and trapped ion mobility enables a rapid, informative method for native monoclonal antibody and antibody-drug conjugate charge variant analysis. This poster was a collaboration with Bruker and NIH and presented at ASMS 2023 as poster WP040.
Fast Screening and Characterization of Therapeutic Peptide by Online Capillary Electrophoresis – Mass Spectrometry (CE-MS)
Presenting a novel strategy for characterization of therapeutic peptides was developed based on microchip CE-MS and linkage-specific derivatization. This poster was presented in collaboration with Ningbo University at ASMS 2023 as poster TP544.
Continuing the Investigation of Microchip Capillary Electrophoresis Coupled with Mass Spectrometry in the Bottom-Up Characterization of Progressively Larger RNAs
Exploring the merits of capillary-zone electrophoresis (CZE)-mass spectrometry to analyze digestion mixtures of large RNAs and attain sequence information. This poster was presented as a collaboration with UConn and Bruker at ASMS 2023 and was poster TP532.
Determination of ATP, ADP, AMP, and Adenosine Levels by Microchip Capillary Electrophoresis Coupled with High Resolution Mass Spectrometry
In this study, we present a novel ZipChip coupled with a QEHF for adenosine nucleotide analysis. This poster was presented at ASMS 2023 as poster TP489.
Sample to Result Workflow for the Investigation of Biosimilars vs Innovator Cetuximab by Charge Variant Analysis using Microchip CE-MS
This poster was presented in collaboration with Protein Metrics and highlights how the ZipChip coupled to HRMS provides a streamlined analytical approach to rapidly assess the heterogeneity of the investigated therapeutic proteins through the comprehensive interrogation of the different charge variants and their intact MS spectra. This poster was presented at ASMS 2023 as poster TP 384.
Peptide Mapping Workflow for Direct Microchip CE-MS Analysis of Biopharmaceuticals
In this study we demonstrated the suitability of a modified digestion workflow for peptide mapping analysis via CE separation. This poster was presented at ASMS 2023 in collaboration with NIBRT. It was poster MP619.
Combining Capillary Electrophoresis and Trapped Ion Mobility Spectrometry Mass Spectrometry to Analyze Epitranscriptomic Marks Mediating Virus-Host Interactions in Infectious Diseases
This poster evaluates the merits of capillary-zone electrophoresis (CZE)-MS. This poster was presented at ASMS 2023 as a collaboration with UCONN. It was poster MP548.
Use of Microchip Capillary Electrophoresis – Mass Spectrometry for Automated Rapid Measurement of Enzyme Reaction Kinetics
This approach allows rapid measurement of enzymatic cleavage dynamics in real time. It can be applied to almost any enzyme substrate reaction and is highly tolerable to various buffers, thanks to the CE Chip-MS configuration. This poster was presented at ASMS 2023 with collaborators as poster MP547.
Rapid CE-MS Analysis of Released N-Glycan: Optimized Workflow for Direct CE Compatibility
N-glycan profiling is a critical step of biopharmaceuticals analysis as N-glycans play an important part in both activity and safety of monoclonal antibodies and other glycosylated biotherapeutics. This poster was presented at ASMS 2023 as poster MP291.
In-depth Metabolic Characterization at Different Phases of Cell Culture with a Turnkey CE-ESI-HRMS Workflow
Learn about a turnkey CE-MS metabolomics workflow solution used to characterize media metabolites and intracellular metabolites of CHO bioreactors. The poster was on display at Bioprocessing Summit, and ESACT #253 in 2024.
Innovations in Analytical Technology for Cell & Gene Therapy Manufacturing
This poster highlights our at-, on-, and in-line analytical solutions that contribute to an enhanced understanding, characterization, and control of bioprocesses in the development and manufacturing of cell and gene therapies.
Optimized Native Conditions for Charge Variant Analysis Using a Microfluidic CE-MS Platform Yield Higher Quality Data and Enables Analysis of Lower Titer Samples
Learn about the improvements made in the new Charge Variant Analysis kit, compared to the previous Intact Antibodies and Native Antibodies kits. ASMS 2024.
Charge Variant Analysis of IgGs Directly from Cell Culture Supernatant Using Microfluidic CE-ESI-MS
This poster describes the application of Microfluidic CE-ESI-MS for analyzing protein charge variants directly from a bioreactor. It investigates the feasibility of a “dilute and shoot” approach compared to a buffer exchange method for sample preparation. ASMS 2024.
Raman-based Lactate Feeding Impact on Culture Characteristics and Product Quality in Mammalian Cell Cultures
This research explores Raman spectroscopy for real-time glucose monitoring and feeding strategies in CHO cell cultures, aiming to improve product quality.
Improving Cell and Gene Therapy Manufacturing Processes by Automated On-line and In-line Bioprocess Analytical Technologies
Recent advances in cell and gene therapy bioprocess analytical technology regimes, approaches to de novo analytical modeling/calibration of these systems, and how their informing power can be leveraged with the increasing sophistication of bioprocess metabolic models for predictive bioprocess optimization. View our poster from the 2024 Advanced Manufacturing Cell and Gene Therapies Conference. This was poster 29. This poster was also on display as #843 at ISCT 2024.
Bottom-Up Analysis of Progressively Larger RNAs by CZE-MS and CZE-MS/MS
In this poster from ASMS 2022, researchers from University of Connecticut and Bruker showed that capillary zone electrophoresis performed on a chip proved to be an effective alternative to liquid chromatography as a front-end separation system to analyze both pure samples and complex digestion mixtures of nucleic acids.
Multi-level Expeditious Characterization of Adeno-associated Virus (AAV) Capsid Proteins Using ZipChip Microfluidic CE Coupled with HRMS
In this poster from ASMS 2022, scientists from Thermo Fisher Scientific and 908 Devices present a novel approach using microchip CE-MS for the characterization of AAVs.
A Sensitive and High-Throughput Assay for Analyzing Peptide Exchanged MHCl by High-Resolution Native CZE-MS
In this poster from ASMS 2022, researchers from Genentech describe a sensitive and high-throughput assay using capillary-zone electrophoresis coupled to a high-resolution mass spectrometer for characterizing peptide-exchanged MHCI under native conditions.
Combining μSPE ZipChip CE with PRM-LIVE on a timsTOF Pro for Warp-speed Selectivity Profiling of Deubiquitinase (DUB) Small Molecule Inhibitors
In this ASMS 2022 poster, researchers from Dana Farber Cancer Institute and Harvard Medical School developed a ZipChip-based method for facilitating high-throughput identification
of new small molecule inhibitors.
Microchip CE-MS Analysis of Sialylated N-glycan Linkage Isomers
In this ASMS 2022 poster, Fudan University researchers show a ZipChip-based strategy for qualitative and quantitative analysis of sialylated N-glycan linkage isomers was developed.
CE-MS of Oligonucleotides using a ZipChip
This ASMS 2022 poster from Boehringer Ingelheim explores the use of ZipChip coupled to a high resolution mass spectrometer as an orthogonal method to chromatography for oligonucleotide analysis.
Rapid Characterization of Oligos and Related Impurities Using Microchip CE-MS without Ion-pairing Reagents
This poster from ASMS 2022 describes a simple & fast method for oligonucleotide characterizations using a Microchip CE-MS platform.
Glycosylation Profiling of mAB Subunits with CZE-MS
This ASMS 2022 poster from Bruker shows ZipChip coupled with high resolution mass spectrometry providing charge-based selectivity which can assist with glycoforms separation.
Pushing the Boundaries of Speed and Sensitivity in Proteomics
To overcome sample loading limitations in CE-MS, the poster authors engineered a 1 mm SPE bed within the chip geometry, allowing 100-fold improvements in concentration sensitivity limit for CE-MS.
A Broad Application of Migration Time Indexing and Metabolite Libraries to Evaluate Quantitative Coverage in Microchip CE-MS Metabolomics
In the field of metabolomics, microchip capillary electrophoresis (mCE) has a number of potential advantages over traditional liquid chromatography (LC), including higher throughput due to decreased separation times, increased separation efficiency, and lower required sample volumes.
A Microchip CE-MS Based End-to-End Solution for the Analysis of Synthetic Oligonucleotides
This poster showcases a workflow using microchip CE-MS that is coupled with a vendor-neutral informatics platform
as a complete solution for the analysis of synthetic oligos.
Microchip CE-MS with Integrated Solid Phase Extraction
Microchip CE-MS of Oligonucleotides: What Can It Separate?
Fast Chip-CEMS for Characterization of Protein/PTM Changes
Combined Top-down and Bottom-up Proteomics
ZipChip for Glycomics and Glycoproteomics
Fast and Efficient Biopharmaceutical Characterization
Characterization of charge heterogeneity of mAb
Rapid, Multi-level Analysis of Complex Proteins
Oligonucleotide Analysis via Microfluidic CE-MS
Advancing mAb Characterization with Microchip CE-MS Couples to a PASEF Enabled QTOF
96 Bottles of Beer: Metabolic Profiling of Spent Growth Media
Native Analysis of Monoclonal Antibodies
Characterization of Protein/PTM Changes
Native CE-MS for Biopharmaceutical Characterization
A Rapid Microfluidic Method for Molecular Weight Determination and Spent Media Analysis of an IgG1 Intact Protein in Growth Media
Bioprocessing of Monoclonal Antibody Based Therapies
NIBRT Presents Native MS for Charge Variants using ZipChip
NIBRT Presents ZipChip for Native Charge Heterogeneity
Monitoring Amino Acid Composition of Cell Culture Media
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Published Articles
Development of a rapid and highly accurate method for 13C tracer-based metabolomics and its application on a hydrogenotrophic methanogen
Yuto Fukuyama, et al. ISME Communications, 2024.
The novel 13C tracer-based metabolomics method using ZipChip CE-MS provides highly accurate and sensitive identification of central carbon metabolism and amino acid biosynthesis pathways, enhancing our understanding of microbial metabolic capabilities and revealing previously unrecognized alternative enzymes.
The Critical Role of CE-MS Methods for Biotherapeutic Characterization
Capillary electrophoresis (CE) methods play an important role in biomolecular characterization while microfluidic CE-mass spectrometry (MS) accelerates analytical workflows and improves development/commercialization timelines.
Identification of AAV capsid proteins
Characterization of a SRPK1/2 Inhibitor Complex
Detection and Quantification of neurotoxin β-N-methylamino-l-alanine
Characterizing Covalent CDK Inhibitor Complexes
Rapid Fingerprinting of a Highly Glycosylated Fusion Protein
Analysis of monosaccharides, oligosaccharides and glycopeptides
Metabolite Profiling Reveals Predictive Biomarkers
Monitoring Glycosylation Profile and Protein Titer in Cell Culture Samples
Characterizing Antiviral Protein-Inhibitor Complexes
Characterization of NIST mAb Proteoforms and Glycoforms
In-depth analysis of monoclonal antibodies using microfluidic capillary electrophoresis and native mass spectrometry
Comparative Elucidation of Cetuximab Heterogeneity on the Intact Protein Level
Characterization of Protein Assemblies Related to Cancer Proliferation
Analysis of alkaloids in plant cell cultures
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Download this Safety Data Sheet (SDS) for documentation related to the properties of each chemical in the product; the physical, health, and environmental health hazards; protective measures; and safety precautions for handling, storing, and transporting the product.
Deoxyribonucleic Acid, Unmodified
SDS Deoxyribonucleic Acid, Unmodified
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SDS Standard: Amino Acids (EU)
SDS Standard: Amino Acids (UK)
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SDS Additive: Charge Variant Analysis (EU)
SDS Additive: Charge Variant Analysis (UK)
SDS Additive: Charge Variant Analysis (USA)
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BGE: Charge Variant Analysis (EU)
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Deoxyribonucleic Acid, Unmodified
SDS Deoxyribonucleic Acid, Unmodified
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ZipChip Interface
Capillary electrophoresis-mass spectrometry (CE-MS) has never been more convenient. The compact design of the ZipChip Interface seamlessly integrates all fluidics and electronics—just clip it on your mass spec and off you go!
Mass Spectrometer Compatibility Thermo Fisher Scientific: ZCI-Classic
- Exactive: Plus, Plus EMR
- Q Exactive: Biopharma, HF Biopharma, HF-X Biopharma, UHMR, Plus, HF,
- HF-X, Focus
- LTQ: XL, Orbitrap XL
ZC-Ti
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MX Biopharma
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- TSQ: Triple Quad, Altis Triple Quad, Endura Triple Quad SCIEX: ZC-Si
- QTRAP: 4500, 6500+
- Triple Quad: 3500, 4500, 5500, 5500+, 6500, 6500+
- Triple TOF: 5600, 6600, 6600+ BRUKER: ZC-Bi
- Compact
- Impact
- maXis II
- timsTOF: Pro 2, fleX, fleX MALDI-2, HT, MALDI PharmaPulse
- solariX
- scimaX
Specifications Software: - Standalone ZipChip acquisition software
- Integrates with your MS software Dimensions: 7” W x 6.5” H x 11” D /18cm W x 17cm H x 28cm D Weight: 16 lbs / 7.3 Kg Power Requirements: 24V DC, 65W Laser Type: Class 1 Laser Product*
*This equipment contains a 520 nm, 4.5 mW Class 3R laser diode with a beam divergence of 1.5 mrad but is classified as a Class 1 Laser Product as an end-use as evaluated to IEC 60825-1:2014. Avoid direct eye exposure to the beam and do not remove the laser for any other purpose.
Autosampler*
Achieve unattended sample runs from samples vials or 96-well plates. The platform allows you to choreograph sample runs using ZipChip software.
Specifications Sample Format: Vials or standard well plates (47 mm max height) Sample Capacity: Two sample trays that hold 48 vial racks or 96-well plates Sample Tray Cooling: - Built-in Peltier cooling
- Minimum: 4°C ± 2°C
- Maximum: ambient temperature –3°C
(Measured as air temperature in sample compartment for maximum air temperature 25°C and maximum humidity 80%) Dimensions: 11.75” W x 14.25” H x 22” D /30cm W x 36cm H x 57.5cm D Weight: 46.3 lbs / 21 kg Power Requirements: 100–240 VAC +/-10%, 50–60Hz, 200 VA
*Required for ZipChip Interface, Autosampler version
[post_title] => ZipChip Hardware Specification Sheet
[post_excerpt] => The compact interface seamlessly integrates all fluidics and electronics. Just clip it on your mass spec and off you go!
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ZipChip Interface
Capillary electrophoresis-mass spectrometry (CE-MS) has never been more convenient. The compact design of the ZipChip Interface seamlessly integrates all fluidics and electronics—just clip it on your mass spec and off you go!
Mass Spectrometer Compatibility Thermo Fisher Scientific: ZCI-Classic
- Exactive: Plus, Plus EMR
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- HF-X, Focus
- LTQ: XL, Orbitrap XL
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- Compact
- Impact
- maXis II
- timsTOF: Pro 2, fleX, fleX MALDI-2, HT, MALDI PharmaPulse
- solariX
- scimaX
Specifications Software: - Standalone ZipChip acquisition software
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*This equipment contains a 520 nm, 4.5 mW Class 3R laser diode with a beam divergence of 1.5 mrad but is classified as a Class 1 Laser Product as an end-use as evaluated to IEC 60825-1:2014. Avoid direct eye exposure to the beam and do not remove the laser for any other purpose.
Autosampler*
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Specifications Sample Format: Vials or standard well plates (47 mm max height) Sample Capacity: Two sample trays that hold 48 vial racks or 96-well plates Sample Tray Cooling: - Built-in Peltier cooling
- Minimum: 4°C ± 2°C
- Maximum: ambient temperature –3°C
(Measured as air temperature in sample compartment for maximum air temperature 25°C and maximum humidity 80%) Dimensions: 11.75” W x 14.25” H x 22” D /30cm W x 36cm H x 57.5cm D Weight: 46.3 lbs / 21 kg Power Requirements: 100–240 VAC +/-10%, 50–60Hz, 200 VA
*Required for ZipChip Interface, Autosampler version
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Tech Notes
How It Works
Download this ZipChip Tech Note to learn how the system works.
Performance Validation Study
Using the ZipChip Infusion Function
Using Field Strength Gradients
ZipChip and Detergents
Evaluation of Sample Carryover
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Testimonials
ZipChip Customer Experience: NIBRT – 908 Devices Exceeds in Support
Dr. Josh Smith from NIBRT discusses the incredible customer support he receives from 908 Devices.
ZipChip Customer Experience: University of Wisconsin – ZipChip is easy to use
Ye Ge from the University of Wisconsin discusses how easy ZipChip is to use, with fast separations, and minimal training.
ZipChip Customer Experiences
Hear from our various experts on why they use ZipChip in their lab.
ZipChip Customer Experience: NIBRT – Unique ZipChip Advantages
Hear from Dr. Sara Carillo from NIBRT on the unique advantages of ZipChip for charge variant analysis.
ZipChip Customer Experience: University of Wisconsin – Speed up Analysis for Complex Biological Matrices
Hear from Li Lingjun from the University of Wisconsin on how ZipChip’s separation techniques speed up analysis and simplify workflows in the academic lab.
ZipChip Customer Experience: Symphogen – Charge Variant Separation
Listen to Dan Bach Kristensen, a principal scientist at Symphogen, as he shares how ZipChip helps him with charge variant separation.
Interview: The Key to Unsolved Insights into RNA Characterization?
Recorded at ASMS 2022, Dan Fabris from the University of Connecticut shares his experience with RNA characterization using ZipChip.
Interview: Oligonucleotide Analysis with ZipChip at Boehringer Ingelheim
Recorded at ASMS 2022, Vladimir Papov shares his experience with oligo analysis using ZipChip.
Hear from Our Users: The Need for Speed in the Lab
Listen to ZipChip and REBEL users from The Dana Farber Cancer Institute, Sartorius, and Amgen discuss the impact of faster workflows in the lab.
Testimonial – Zachary Kelley – University of Kentucky
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RSK Media, Rizwan Chaudhrey, interviews 908 Devices CEO and co-founder, Kevin J. Knopp. This interview highlights:
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RSK Media, Rizwan Chaudhrey, interviews 908 Devices CEO and co-founder, Kevin J. Knopp. This interview highlights:
- A background on 908 Devices and why they started the company.
- What the key drivers are behind 908 Devices product launches.
- Why 908 Devices acquired RedWave Technology.
- Kevin J. Knopp provides his thoughts on the future for Life Science.
- Explains what differentiates 908 Devices from other companies.
[post_title] => Interview: RSK Media Interviews Kevin J. Knopp
[post_excerpt] => RSK Media, Rizwan Chaudhrey, visited Boston and met with 908 Devices CEO, Kevin J. Knopp. Topics included key business drivers, the RedWave Technology acquisition, and the future of life sciences.
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Videos
Interview: RSK Media Interviews Kevin J. Knopp
RSK Media, Rizwan Chaudhrey, visited Boston and met with 908 Devices CEO, Kevin J. Knopp. Topics included key business drivers, the RedWave Technology acquisition, and the future of life sciences.
Whiteboard Explainer: Avoiding the Stickiness of Oligo Analysis
If you have worked with oligos before, you know this can be a challenging application. Fortunately, ZipChip offers a way to make workflows faster and easier.
ZipChip. Put your mass into it
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About this Webinar
Understanding how bioprocess changes impact product quality is crucial for successful biopharmaceutical development. However, frequent data collection across the entire bioprocess can be a significant challenge for analytical teams.
This webinar explores a streamlined solution for acquiring critical mAb quality attributes, including charge variant profiles, sequence identity, glycosylation profiles, and glycation levels. We will demonstrate a simple "dilute-and-shoot" analysis utilizing ZipChip capillary electrophoresis coupled with high-resolution mass spectrometry.
Furthermore, we will present a case study comparing CHO fed-batch processes with traditional daily bolus glucose feeding and a 2g/L dynamic glucose feed strategy, enabled by MAVEN monitoring and control. This study highlights the clear impact of process feeding strategies on product quality profiles.
Presenters
- Erin Redman, PhD, Principal Scientist, 908 Devices
- Graziella Piras, PhD, Senior Director of Strategic Marketing, Life Science, 908 Devices
Key Takeaways:
- Streamline mAb product quality assessment throughout bioprocess runs.
- Gain insights into charge variant profiles, sequence identity, glycosylation profiles, and glycation levels.
- Understand the link between process feeding strategies and product quality.
- Learn how bioprocess monitoring enables control, such as stabilizing glucose levels at 2g/L via MAVEN control to reduce glycation.
- Explore the benefits of both off-line analytical and on-line process analytical technology (PAT) solutions.
- Discover the efficiency of the dilute-and-shoot ZipChip-MS CVA workflow for frequent measurements.
[post_title] => Streamlining mAb Quality Assessment Throughout the Bioprocess with ZipChip-MS
[post_excerpt] => Dilute-and-shoot ZipChip-MS CVA workflow enabling frequent measurements.
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About this Webinar
A broad range of separation techniques are now routinely combined with mass spectrometry for in-depth biopharmaceutical characterization. Simultaneously, MS software tools have evolved to support the storage, processing, and reporting of the expanding MS data oceans. Here recent progress, challenges, and learnings from MS-based workflows of biopharmaceutical charge variants will be presented and illustrated through case-studies.
In this webinar, you will learn:
- The crucial role that CE-MS plays in a biopharma development lab for the comprehensive characterization of biopharmaceuticals.
- How CE-MS can be used to improve the efficiency of characterization workflows by eliminating method development and increasing the throughput while gaining deeper insights through in-depth analysis of biopharmaceuticals.
Who should attend:
- Scientists working in protein characterization labs for biotherapeutics
- Analytical scientists working in biopharmaceutical labs
- CDMO labs looking to optimize workflows and improve efficiency
Improve Process Intensification
Process intensification requires a thorough understanding of the process to achieve desired yields while improving resource utilization and maintaining consistent product quality. Continuous monitoring of glucose and lactate provides deeper process understanding. Dynamic glucose control can lead to better cell viability, lower levels of lactate production, and therefore, higher titer and better control of quality attributes (CQAs) in fed-batch and intensified processes. Learn how you can make more informed decisions with in-line, on-line and at-line analytics of key process parameters
About this Webinar
Peptide mapping is considered the gold standard for the analysis of biopharmaceuticals. Classical analysis is performed through long workflows involving extensive sample handling and long liquid chromatography (LC-MS) analysis times.
The distinct advantages of short analysis times and unique separation mechanisms makes Capillary Electrophoresis (CE-MS) an attractive alternative for the peptide mapping analysis. Nevertheless, most existing sample preparation protocols are either incompatible with CE-MS analysis or require additional steps for salt removal that may increase sample handling.
Key Takeaways
- An overview of the ZipChip Microchip CE-MS technology and its typical application space
- How to optimize CE-MS compatible sample preparation using a total workflow length of 2 hours
- Data analysis will provide full sequence coverage and accurate quantitation of product quality attributes
- Wide applicability of the workflow to different IgG subclasses will be demonstrated
- A panel discussion (Q&A) at the end
- Microchip CE-MS technology, its capabilities, and typical application space
- How CE-MS technology can enhance peptide mapping throughput
- Coupling peptide mapping sample preparation of monoclonal antibodies to CE-MS analysis
- CE-MS analysis provides robust determination of quality attributes in less than 15 minutes
Meet the Speakers
Sara Carillo
Dr. Sara Carillo completed her Ph.D. in Chemical Sciences in 2013 at the University of Naples “Federico II”. Under the guidance of Prof. Corsaro, she focused on the structural characterisation of polysaccharides and glyco-conjugates from Gram-negative bacteria via NMR and mass spectrometry techniques, focusing on the immunological properties and potentials of extremophiles endotoxins. After a period at the University College of Dublin, in 2015 she joined Dr. Jonathan Bones’s research group in NIBRT working on understanding of the effects of extractables and leachables from single-use bioreactors on CHO cells N-glycome and produced monoclonal antibodies. She is now working in NIBRT as Bioanalytical Research Lead and is mainly interested in the development of new analytical approaches to ensure a deeper and easier understanding of biomanufacturing processes and biopharmaceuticals’ structural complexity.
Adi Kulkarni
Adi Kulkarni is a Principal Scientist. Adi has extensive experience in applying CE-MS in biopharma characterization workflows using different mass spec platforms. He has developed novel applications based on the ZipChip technology and managing strategic collaborations and partnerships. Prior to working at 908 Devices, Adi was an applications scientist at Bruker Daltonics where he worked on Bruker’s Q-TOF line of products. He received his Ph.D. at the University of Massachusetts in 2011 where he developed novel methodologies for the synthesis of nitrogen-containing heterocycles. After his Ph.D., Adi was a postdoctoral fellow at University of Southern California’s Loker Hydrocarbon Research Institute where he discovered new reagents for applications in organofluorine chemistry. He has authored 18 peer-reviewed articles.
Jonathan Bones
Jonathan Bones is the principal investigator of the Characterisation and Comparability Laboratory at NIBRT. An analytical chemist by training, Jonathan’s research group specializes in the development and application of analytical solutions for problems associated with the manufacture and characterization of biopharmaceuticals. This includes recombinant proteins, monoclonal antibodies, and advanced therapies like AAV based gene therapy, cell therapy, and mRNA using their core excellence in analytical separations coupled to mass spectrometry.
Erin Redman
Erin Redman is a Principal Scientist in the Research and Development division of 908 Devices. She joined the company in 2016 and has worked to develop protein characterization assays using the ZipChip technology and further the innovation microfluidic CE separations coupled to mass spectrometry. Prior to starting her career at 908 Devices, she received her Ph.D. under the direction of Professor J. Michael Ramsey at the University of North Carolina at Chapel Hill. There she developed microfluidic technology for interfacing high resolution separations with electrospray mass spectrometry for the characterization of intact biotherapeutic proteins and proteins from biofluids.
About this Webinar
Understanding how bioprocess changes impact product quality is crucial for successful biopharmaceutical development. However, frequent data collection across the entire bioprocess can be a significant challenge for analytical teams.
This webinar explores a streamlined solution for acquiring critical mAb quality attributes, including charge variant profiles, sequence identity, glycosylation profiles, and glycation levels. We will demonstrate a simple "dilute-and-shoot" analysis utilizing ZipChip capillary electrophoresis coupled with high-resolution mass spectrometry.
Furthermore, we will present a case study comparing CHO fed-batch processes with traditional daily bolus glucose feeding and a 2g/L dynamic glucose feed strategy, enabled by MAVEN monitoring and control. This study highlights the clear impact of process feeding strategies on product quality profiles.
Presenters
- Erin Redman, PhD, Principal Scientist, 908 Devices
- Graziella Piras, PhD, Senior Director of Strategic Marketing, Life Science, 908 Devices
Key Takeaways:
- Streamline mAb product quality assessment throughout bioprocess runs.
- Gain insights into charge variant profiles, sequence identity, glycosylation profiles, and glycation levels.
- Understand the link between process feeding strategies and product quality.
- Learn how bioprocess monitoring enables control, such as stabilizing glucose levels at 2g/L via MAVEN control to reduce glycation.
- Explore the benefits of both off-line analytical and on-line process analytical technology (PAT) solutions.
- Discover the efficiency of the dilute-and-shoot ZipChip-MS CVA workflow for frequent measurements.
Webinars On Demand
Streamlining mAb Quality Assessment Throughout the Bioprocess with ZipChip-MS
Dilute-and-shoot ZipChip-MS CVA workflow enabling frequent measurements.
Charge Variant Analysis for Intact Protein Characterization in a Biopharmaceutical Development Lab – Lessons Learned and the Road Ahead
How CE-MS can be used to improve the efficiency of characterization workflows by eliminating method development and increasing the throughput while gaining deeper insights through in-depth analysis of biopharmaceuticals.
Data-driven Process Intensification – Utilizing Smart Analytics to Achieve your Bioprocess Goals
This BPI presentation discusses process intensification and rapid analytics of critical parameters for cell culture, which is necessary to control the process and end product. See how fast and simple analytics significantly speed up the development and optimization processes.
Rapid Clone Screening of Biosimilar Candidates Using Microfluidic CE-MS
In this GEN webinar, our expert speakers, Erin Redman, PhD, and Ruben Cageling, will describe how the ZipChip microfluidic CE-MS technology enables rapid early-stage clone screening for biotherapeutics development. They will present a case study where the ZipChip was used to select clonal cell lines based on their charge variants and glycosylation profile. Furthermore, they will also highlight the new CVA kit, which enabled better separation and sensitivity in charge variant analysis of biotherapeutics.
Direct Microchip CE/MS Peptide Mapping Workflow for the Analysis of mAb CQAs
Microchip CE-MS technology, its capabilities, and typical application space.
EPO Separation & Analysis
Straightforward separation and analysis of a highly glycosylated protein
Simplifying Oligonucleotide Characterization with Microchip CE-MS
In this on-demand webinar, we present a microfluidic capillary zone electrophoresis-based separation coupled with HRAM mass-spectrometry for rapid characterization of oligos in positive ionisation mode.
Analysis of inhibitor-protein complexes and small molecules
Fast Therapeutic Protein Characterization
In-depth characterization of complex monoclonal antibodies and antibody-drug conjugates with native ZipChip CE-MS
Rapid, comprehensive biotherapeutics characterization
NIBRT presents ZipChip for biopharma characterization
In this webinar NIBRT demonstrates ZipChip’s ease of use and flexibility from the control of upstream processes to downstream biotherapeutics characterisation.
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