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Mass spectrometry analysis of in vitro nitration of a recombinant human IgG1 monoclonal antibody
Author(s) -
Liu Hongcheng,
GazaBulseco Georgeen,
Chumsae Chris,
Radziejewski Czeslaw H.
Publication year - 2007
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.3322
Subject(s) - nitration , chemistry , tetranitromethane , tyrosine , residue (chemistry) , monoclonal antibody , recombinant dna , mass spectrometry , chromatography , antibody , biochemistry , organic chemistry , biology , gene , immunology
Nitration of a recombinant human monoclonal antibody was carried out in vitro by incubating the antibody with the nitrating reagent tetranitromethane (TNM). The susceptible sites of nitration were identified using high‐performance liquid chromatography/mass spectrometry (HPLC/MS). In general, tyrosine residues in the variable domains of the antibody are more susceptible to nitration, while tyrosine residues in the constant domains are relatively resistant to nitration. However, one tyrosine residue in the CH1 domain and one tyrosine residue in the CH2 domain are highly susceptible to nitration. Interestingly, the susceptible tyrosine residue in the CH2 domain is followed by the conserved asparagine residue that is glycosylated. Copyright © 2007 John Wiley & Sons, Ltd.