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Anal Biochem. 2011 Aug 15;415(2):212-4. doi: 10.1016/j.ab.2011.04.030. Epub 2011 Apr 27.

Fast analysis of recombinant monoclonal antibodies using IdeS proteolytic digestion and electrospray mass spectrometry.

Author information

1
Analytical Department, LFB Biotechnology, 3 Avenue des Tropiques, 91958 Courtaboeuf (Les Ulis), France. chevreuxg@lfb.fr

Abstract

We describe a fast and informative method to investigate the posttranslational modifications of monoclonal antibodies (MAbs). The MAb is first digested by a specific enzyme that cleaves heavy chains under the hinge domain. After reduction of disulfide bridges, three polypeptide chains of approximately 25 kDa are released and analyzed by liquid chromatography-mass spectrometry (LC-MS). By bisecting the heavy chains prior to MS analysis, this method provides a better MS resolution and facilitates the study of the N-linked glycans as well as of other modifications (loss of C-terminal lysine, pyroglutamination, and oxidation). The sample preparation and analysis can be performed within few hours.

PMID:
21596014
DOI:
10.1016/j.ab.2011.04.030
[Indexed for MEDLINE]

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