1. Fast separation and analysis of reduced monoclonal antibodies with capillary zone electrophoresis coupled to mass spectrometry.
- Author
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Zhao, Yimeng, Sun, Liangliang, Knierman, Michael D., and Dovichi, Norman J.
- Subjects
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SEPARATION (Technology) , *MONOCLONAL antibodies , *CAPILLARY electrophoresis , *ELECTROPHORESIS , *MASS spectrometry , *IMMUNOGLOBULINS - Abstract
Capillary zone electrophoresis–electrospray ionization–mass spectrometry (CZE–ESI–MS) was used for analysis of reduced antibodies. We first developed a simple protocol to condition commercial linear-polyacrylamide coated capillaries for use in top–down proteomics. We then suspended reduced antibodies in a solution of 35% acetic acid, 50% acetonitrile in water. Heavy and light chains were baseline resolved within 10 min and with 3–30 µg/mL detection limits using a 0.1% aqueous formic acid background electrolyte. Quintuplicate runs of a two-antibody mixture produced relative standard deviations of ∼1% in migration time and 10% in peak amplitudes. Resolution was further improved for the two-antibody mixture by using 5% acetic acid as the background electrolyte, highlighting the potential of capillary electrophoresis–mass spectrometry for analysis of antibody mixtures. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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