Proteins and antibodies, whether naturally occurring or manufactured, exist as a population of variants. The multibillion dollar BioPharma industry continues to expand, however for biomolecules to be used as therapeutic agents, they must be rigorously characterized to ensure safety, efficacy, and potency. Due to their complex make-up, attributable to structural modifications during cell culture production, purification, and storage, analysis is extremely challenging and time consuming.

Our recently launched ZipChip™, is an innovative, front-end system that provides high-quality separation capabilities for traditional mass spectrometers. ZipChip has been getting a lot of attention from the industry and academics as it enables full characterization of intact proteins and antibody variants in less than three minutes. A recently published paper from our science founder, Dr. J. Michael Ramsey, and his team at the University of North Carolina at Chapel Hill, looks at monoclonal antibodies (mAbs) as an example.

Traditionally, scientists have leveraged chromatographic and electrophoresis techniques with non-selective detectors to analyze these materials, however these techniques only provide complementary information to the overall characterization of an intact protein or antibody. Mass Spectrometry (MS) has proved to be an ideal technique due to its unique identification capabilities, however pre-separation dramatically simplifies the data for analysis. Unfortunately, most historical, ‘stand-alone’ separation techniques that work for proteins are not directly compatible with MS instruments as they require high salt content.  Without an MS detector these techniques lack the specificity to positively identify individual variants.

When it comes to analyzing intact proteins and antibodies, ZipChip is a real game changer for scientists, as it is directly MS-compatible and separates these variants with only small amounts of sample and in about three minutes of analysis time. For this process, scientists simply load the sample onto the ZipChip HR chip, which was specifically optimized for intact large molecule analysis. ZipChip uses integrated microfluidic technology to prepare, separate by capillary electrophoresis (CE), and then electrospray (ESI) biological samples directly into traditional MS instruments. Since the process is relatively gentle, MS results typically show ‘near native-state’ protein variants with a full mass spectrum behind every electropherogram peak.

With ZipChip, we are able to offer scientists a rapid and generic strategy for the separation of intact proteins and antibodies that facilitates the identification of variants through MS detection. For more information about how this technology has been applied to intact proteins and antibodies, we invite you to read this recently published technical paper in Analytical Chemistry titled “Integrated Microfluidic Capillary Electrophoresis-Electrospray Ionization Devices with Online MS Detection for the Separation and Characterization of Intact Monoclonal Antibody Variants.” This piece is the first demonstration of intact mAb charge variant separations with CE-ESI-MS detection.

Chris Petty, VP Business Development and Marketing