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Identification of N‐terminal modification for recombinant monoclonal antibody light chain using partial reduction and quadrupole time‐of‐flight mass spectrometry
Author(s) -
Yu Lei,
Remmele Richard L.,
He Bing
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2790
Subject(s) - chemistry , monoclonal antibody , recombinant dna , mass spectrometry , chromatography , immunoglobulin light chain , peptide , electrospray ionization , quadrupole time of flight , electrospray , time of flight mass spectrometry , biochemistry , antibody , ionization , organic chemistry , ion , gene , biology , immunology
Since most recombinant monoclonal antibodies (mAbs) contain glutamic acid or glutamate at their N‐terminus, cyclization of these residues to form pyroglutamate is an important degradation pathway that often occurs in therapeutic mAb development. In this work, a rapid method was developed to determine pyroglutamate at the N‐terminus of mAb light chain by liquid chromatography coupled with electrospray ionization on a quadrupole time‐of‐flight mass spectrometer (QTOF). High levels of pyroglutamate were found at the N‐terminus of the light chain of a typical recombinant mAb. The quantitative results were comparable to those obtained with a more conventional peptide mapping method. The direct method outlined here can be used to evaluate the impact of N‐terminal cyclization during the processing of recombinant mAbs. Copyright © 2006 John Wiley & Sons, Ltd.