Interchain Disulfide Bonding in Human IgG2 Antibodies Probed by Site-Directed Mutagenesis

被引：24

作者：

Allen, Martin J. ^{[1
]}

Guo, Amy ^{[1
]}

Martinez, Theresa ^{[1
]}

Han, Mei ^{[1
]}

Flynn, Gregory C. ^{[1
]}

Wypych, Jette ^{[1
]}

Liu, Yaoqing Diana ^{[1
]}

Shen, Wenyan D. ^{[1
]}

Dillon, Thomas M. ^{[1
]}

Vezina, Christopher ^{[1
]}

Balland, Alain ^{[1
]}

机构：

[1] Amgen Inc, Proc & Prod Dev, Seattle, WA 98119 USA

来源：

BIOCHEMISTRY | 2009年 / 48卷 / 17期

关键词：

FC-GAMMA-RIII; RECOMBINANT PROTEIN EXPRESSION; HUMAN-IMMUNOGLOBULIN G2; STRUCTURAL ISOFORMS; HEAVY CHAIN; VARIANTS; BINDING; ISOTYPE; COMPLEX; FUCOSE;

D O I：

10.1021/bi8022174

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：

Human IgG2 exists as a mixture of disulfide-linked structural isoforms that can show different activities. To probe the contribution of specific cysteine residues to the formation of structural isoforms, we characterized a series of Cys -> Ser mutant IgG2 recombinant monoclonal antibodies, focused on the first C(H)1 cysteine and the first two hinge cysteines. These residues participate in the fort-nation of structural isoforms that have been noted by nonreduced capillary sodium dodecyl sulfate polyacrylamide gel electrophoresis, reversed-phase high-performance liquid chromatography, and cation exchange chromatography. We show that single Cys -> Ser mutants can greatly reduce heterogeneous disulfide bonding in human IgG2 and maintain in vitro activity. The data demonstrate the feasibility of applying site-directed mutagenesis to reduce disulfide bond heterogeneity in human IgG2 while preserving the activity of this therapeutically important class of human antibodies.

引用

页码：3755 / 3766

页数：12

共 34 条

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