Rapid LC-MS screening for IgG Fc modifications and allelic variants in blood

被引：29

作者：

Goetze, Andrew M. ^{[1
]}

Zhang, Zhongqi ^{[1
]}

Liu, Ling ^{[2
]}

Jacobsen, Frederick W. ^{[2
]}

Flynn, Gregory C. ^{[1
]}

机构：

[1] Amgen Inc, Dept Proc & Prod Dev, Thousand Oaks, CA 91320 USA

[2] Amgen Inc, Dept Prot Sci, Thousand Oaks, CA 91320 USA

来源：

MOLECULAR IMMUNOLOGY | 2011年 / 49卷 / 1-2期

关键词：

Allotype; Allele; Fucosylation; Glycation; Fc; Screening; DEPENDENT CELLULAR CYTOTOXICITY; N-LINKED OLIGOSACCHARIDE; IMMUNOGLOBULIN ALLOTYPES; SERUM IGG; STREPTOCOCCUS-PYOGENES; ANTIBODY-RESPONSES; GAMMA-RIIIA; RHEUMATOID-ARTHRITIS; STRUCTURAL-CHANGES; BINDING-AFFINITY;

D O I：

10.1016/j.molimm.2011.09.002

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new method for simultaneously screening allelic variants and certain Fc modifications on endogenous human IgG1 and IgG2 directly from blood samples is described. The IdeS endoproteinase was used to cleave IgG in serum to generate Fe, which, after denaturation, was analyzed directly as monomeric Fe (Fc/2) by LC-MS to identify the haplotype(s) present in each individual. The relative levels of IgG isotype and haplotype ratios were generated along with the profile of the major Fe glycans and several other modifications associated with each IgG1 or IgG2 haplotype. Since only minute quantities (5 mu L) of blood are required and analysis can be highly automated, this approach lends itself to screening large populations. We demonstrate its utility in examining possible correlations between Fc properties and allelic variants. IgG1 core fucosylation, which significantly impacts antibody dependent cellular cytotoxicity (ADCC), showed an asymmetric distribution, with a small number of individuals showing unexpectedly high core afucosylation levels. In these individuals, IgG2 afucosylation levels were normal. Finally, a new IgG1 allotype, previously not characterized, was identified using this analytical methodology. (C) 2011 Elsevier Ltd. All rights reserved.

引用

页码：338 / 352

页数：15

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