Qualitative and semiquantitative analysis of composite mixtures of antibodies by native mass spectrometry
| Autor: | Alexander Berthold Hendrik Bakker, Linda Johanna Aleida Hendriks, Esther van Duijn, Sara Rosati, John de Kruif, Arjan Barendregt, Natalie J. Thompson, Mark Throsby, Albert J. R. Heck |
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| Rok vydání: | 2012 |
| Předmět: |
Bispecific antibody
Spectrometry Mass Electrospray Ionization Chromatography biology Chemistry Ion chromatography Analytical chemistry Peak fitting CHO Cells Mass spectrometry Chromatography Ion Exchange Antibodies Analytical Chemistry Cricetulus Cricetinae Immunoglobulin G Mass spectrum biology.protein Animals Antibody Semi quantitative |
| Zdroj: | Analytical chemistry. 84(16) |
| ISSN: | 1520-6882 |
| Popis: | Native mass spectrometry was evaluated for the qualitative and semiquantitative analysis of composite mixtures of antibodies representing biopharmaceutical products coexpressed from single cells. We show that by using automated peak fitting of the ion signals in the native mass spectra, we can quantify the relative abundance of each of the antibodies present in mixtures, with an average accuracy of 3%, comparable to a cation exchange chromatography based approach performed in parallel. Moreover, using native mass spectrometry we were able to identify, separate, and quantify 9 antibodies present in a complex mixture of 10 antibodies, whereas this complexity could not be unraveled by cation exchange chromatography. Native mass spectrometry presents a valuable alternative to existing analytical methods for qualitative and semiquantitative profiling of biopharmaceutical products. It provides both the identity of each species in a mixture by mass determination and the relative abundance through comparison of relative ion signal intensities. Native mass spectrometry is a particularly effective tool for characterization of heterogeneous biopharmaceutical products such as bispecific antibodies and antibody mixtures. |
| Databáze: | OpenAIRE |
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