Biologics
Optimized Production of Recombinant Proteins
Gain data-driven insights into critical process parameters and product quality attributes to increase product yield, improve process robustness and product quality.
Innovation for Recombinant Protein Manufacturing
The pharmaceutical industry has witnessed significant growth, largely due to the rising demand for biologics—therapeutics such as monoclonal antibodies, hormones, and vaccines that have revolutionized modern medicine. These biologics offer targeted treatments for various conditions, including cancer, autoimmune disorders, and infectious diseases. Recombinant protein drugs, such as monoclonal antibodies (mAbs), stand out as a key therapeutic class due to their specificity, efficacy, and versatility. These genetically engineered proteins are designed to bind specific antigens, enabling targeted therapeutic interventions.
Over the last decade, the manufacturing of recombinant proteins has undergone significant advancements. This has lead to substantial improvements in efficiency, quality, and cost-effectiveness. These developments have been driven by several key technologies and innovations in cell line engineering, media formulations, and process intensification. Additionally, Process Analytical Technology (PAT) and real-time monitoring have enabled precise control over critical process parameters, helping ensure consistent product quality and reduce variability. Despite these advancements, the complex interplay between media components and cellular metabolism, increasing complexity of biologics, coupled with the need for higher productivity and cost-effectiveness, necessitates continuous innovation and improvement in cell culture media formulations and process control strategies.
Optimizing the Upstream Process
Developing and implementing a robust and scalable process can be challenging. Our bioprocessing solutions help you:
- Screen and develop cell culture media optimized to unique needs of the expression system
- Maintain optimal concentrations of nutrients and metabolites in cell culture and fermentation
- Rapidly characterize functional and structural properties of therapeutic proteins
- Gain invaluable data-driven insights to enhance process understanding and model performance
At-line Cell Culture Media Analysis
Identify the optimal concentrations of amino acids in the media to maximize cell growth, viability, productivity, and product quality
Real-Time Glucose Monitoring & Control
Dynamically control glucose feeding to reduce accumulation of growth-inhibiting byproducts and improve quality
On-Line Methanol Monitoring
Monitor methanol in yeast fermentation to optimize induction and prevent excess accumulation
In-line Biomass Trending
Analyze batch growth trajectories with real-time qualitative biomass trending
Protein Characterization
Rapidly assess multiple critical quality attributes of recombinant proteins to ensure product quality and efficacy
Critical Quality Attributes (CQAs) of Biologics are Affected by Nutrients and Metabolites in Cell Culture Media
Finding the Right Balance of Amino Acids in the Media is Critical
Amino acids serve as fundamental building blocks for protein synthesis and are involved in several cellular pathways. Their concentrations in basal media and feeds critically impact the yield, structure, and post-translational modifications (PTMs) of mAbs and recombinant proteins. The levels of specific amino acids like glutamine, cysteine, asparagine, serine, and threonine are particularly influential and amino acid imbalances can cause the accumulation of toxic by-products, which may stress the cells and cause the undesirable PTMs.
- Glutamine is crucial for cell growth and protein production. Its depletion or excess can lead to altered glycosylation patterns, which are critical for the bioactivity and immunogenicity of mAbs.
- Variations in cysteine can affect the formation of disulfide bonds, which are essential for maintaining the structural integrity of the antibody.
- Asparagine levels directly influence the glycosylation of the Fc region of antibodies, impacting their effector functions, including antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC).
- Depletion of serine and threonine, during late-stage cultivation, can result in incomplete glycosylation or improper folding of the proteins, leading to heterogeneity in the product.
REBEL Ensures Media is Optimized for Quality and Yield
Culture media is a critical component of recombinant protein production, providing essential nutrients for cell growth and protein expression. Different clones and cell lines have unique nutrient requirements, necessitating tailored media and feeding strategies. Variability in media composition (vendor-to-vendor and lot-to-lot), caused by small differences in raw materials or media preparation processes, can significantly affect protein yield and quality. Comprehensive analysis of media components is often hindered by core lab turnaround times, slowing process development and decision-making. REBEL simplifies analysis and accelerates actionable intelligence delivery, enabling process development scientists, MSAT, and bioprocessing engineers to make informed decisions efficiently.
Media Screening & Fingerprinting
With REBEL, you can screen and select the best media for your cell line from the start. Different cell lines and clones require specific nutrients for optimal growth and productivity. In process development, media screening allows for the comparison of media formulations between vendors to identify the right balance of nutrients, such as amino acids and vitamins, to support robust cell growth and high cell viability.
Media Development
Developing the optimal media formulation requires frequent monitoring of key nutrients throughout the cell culture process. Waiting for traditional media analysis results can significantly delay process development timelines. REBEL offers a rapid analysis solution, allowing researchers to make timely adjustments to media formulations, improving productivity and accelerating time to market.
Process Modeling
Add the in-process amino acid characterization data to your predictive process models for smarter DOE resulting in faster development, enhanced process understanding, and more optimized culture performance.
Process Monitoring
Perform qualitative media fingerprinting using REBEL to help detect potential mis-formulated media lots and implement routine monitoring of amino acids in production runs to decrease risk of wasted batches and speed up investigations of process deviations.
Accelerate Upstream Process Development with Data-driven Feeding Strategy Optimization
Webinar On Demand Spotlight
Learn how using at-line spent media analysis and on-line glucose and lactate monitoring for the optimization of feeding strategy.
Monitoring and Controlling Critical Process Parameters (CPPs) for Achieving Optimal Cell Culture Conditions
Glucose
Glucose concentration affects the metabolic flux of host cells, balancing energy production, reducing oxidative stress, and optimizing recombinant protein expression. Glucose metabolism also influences amino acid availability by providing precursors and reducing equivalents, crucial for maintaining protein synthesis rates and ensuring proper folding and PTMs. Optimal glucose levels can prolong cell viability and increase yield by preventing nutrient depletion and accumulation of lactate and ammonia. Glucose levels can impact several critical product quality attributes such as glycation and glycosylation, potentially affecting protein structural integrity, function, immunogenicity, and stability.
Lactate
Lactate accumulation can indicate cellular stress and impact cell growth and protein stability. Lactate buildup acidifies the media, affecting protein charge properties and leading to structural and functional changes. High lactate levels can induce cellular stress, causing errors in protein synthesis and compromising sequence accuracy. Continuous lactate monitoring allows timely interventions to maintain optimal culture conditions and preserve protein charge characteristics. By monitoring lactate levels, strategies can be implemented to reduce stress, such as optimizing oxygen supply or adjusting nutrient levels, ensuring accurate protein expression.
Biomass
Tracking biomass trends helps optimize the timing of feeding strategies and harvests. This optimization minimizes waste and maximizes the use of resources, leading to higher productivity. Real-time biomass trending can reveal deviations from expected growth patterns, which may indicate problems such as contamination, nutrient depletion, or metabolic stress. Early detection allows for prompt corrective actions, reducing the risk of batch failures.
Methanol
In yeast fermentation processes, such as those using Pichia pastoris, methanol is often used as an inducer for the expression of recombinant proteins. The concentration of methanol is a key determinant of protein yield and quality. Low methanol concentrations may result in insufficient protein expression, while high concentrations can be toxic to the cells, leading to cell death and reduced productivity. Additionally, methanol metabolism in yeast can lead to the production of formaldehyde and hydrogen peroxide, which can cause oxidative stress, thereby affecting protein folding and leading to the formation of misfolded or aggregated proteins.
Gain Timely Process Insights and Maintain Optimal Concentrations of Nutrients and Metabolites with MAVERICK, MAVEN and TRACE C2
Real-time monitoring and control of critical process parameters are essential for developing robust recombinant protein manufacturing processes. Regardless of the bioreactor type or operation mode (fed-batch or perfusion), continuous monitoring can help achieve key objectives such as maximizing protein yield, optimizing cell growth, and ensuring product quality. Continuous monitoring of critical process parameters provides real-time insights into cellular metabolism, enabling feedback control strategies to automatically adjust nutrient feed rates and prevent nutrient depletion or toxic metabolite accumulation.
Instant Monitoring and Control of Critical Process Parameters (CPPs) with MAVERICK
Data presented in this application note demonstrates how MAVERICK, a Raman spectroscopy-based PAT device, can be used across several CHO and HEK cell lines, eight chemically defined media and bioreactor scales from 250 ml to 10L. MAVERICK can obtain data in minutes without the need for complex chemometric models, thereby eliminating the implementation hurdles inherent to traditional in-line Raman-based PAT process controls.
Ensuring Product Quality and Consistency through Advanced Characterization
Understanding the biotherapeutic profile and verifying heterogeneity, molecular integrity, sequence fidelity, and post-translational modifications during process development are essential for ensuring the safety, efficacy, and consistency of recombinant protein biologics.
- Glycosylation can influence antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC)
- Glycation, non-enzymatic glycosylation on protein amine groups, can block biologically functional sites or further degradation that induces aggregation
- Sequence variants can alter the antigen-binding site (paratope) of the antibody, reducing its affinity and specificity for the target antigen
- Charge heterogeneity can arise from deamidation, glycation, or C-terminal lysine clipping, affecting the antibody’s binding affinity and stability
- Variability in molecular weight may signal issues with structural integrity, such as unintended variants or modifications that could affect its therapeutic function
GlcNAc
Fucose
Galactose
Mannose
Sialic Acid
Conjugate
Rapid Protein Characterization for Quality and Consistency
ZipChip is a cutting-edge microfluidic capillary electrophoresis-mass spectrometry (CE-MS) device for protein characterization workflows. Efficiently separate and seamlessly introduce samples into your mass spectrometer with minimal sample prep and no method development. ZipChip provides rapid, high-resolution analysis across multiple critical quality attributes (CQAs) and post translational modifications. Streamline your analytical workflows and utilize comprehensive and accurate data to support robust development and quality control of recombinant protein therapeutics.
Your Choice for Ultimate Bioprocess Monitoring and Control
Our suite of devices provides the data you need, where and when you need it.
For Cell Culture Media Analysis
REBEL
Analyze fresh or spent media and get quantitative reports for amino acids, dipeptides, water soluble vitamins and biogenic amines.
For Bioprocessing
MAVERICK
Raman-based, in-line monitoring and control of multiple bioprocessing key process parameters in up to six bioreactors simultaneously.
For Glucose & Lactate
MAVEN
Get valuable insights and control from real-time glucose and lactate monitoring without the loss of bioreactor or fermenter volume.
For Methanol or Ethanol
TRACEC2
Monitor methanol or ethanol and automatically control substrate feeding in microbial fermentation.
For Biological Characterization
ZipChip
Rapid protein characterization for efficient, accurate, and comprehensive analysis of multiple CQAs.
Suggested Resources
Identify variations in dry and liquid CHO media
Running a quick fresh media assay prior to feeding the culture ensures consistency of the media components when scaling up or during the transition between liquid-to-dry media blends.
On-line Measurement of Glucose and Lactate with Automated Feedback Control
MAVEN answers the urgent need for on-line measurements and automated control of glucose levels, which reduces manual labor, lowers risk of contamination from sampling, decreases risk of human error with feeding, and reduces the stress that cells may experience from variable glucose levels.
A ZipChip Based CZE-MS Analysis for Quick Assessment of Biotherapeutics
Methanol Dialysis
The online analyzer TRACE C2 Control allows a rapid and precise determination of the methanol concentration in the fermenter and provides the tools for a quick set-up of a feed control strategy
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