Journal
MOLECULAR IMMUNOLOGY
Volume 47, Issue 7-8, Pages 1592-1594Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.molimm.2010.02.021
Keywords
IgG4; Size-exclusion chromatography; HP-SEC; SDS-PAGE; Half-molecule; Immunoglobulin
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Human IgG4 antibodies are dynamic molecules that in vivo exchange half-molecules to become bispecific antibodies. Here we show that IgG4 antibodies and IgG4 Fc fragments similarly exchange resulting in hybrid antibodies (a single Fab + Fc) with a molecular weight of ca. 100 kDa. These antibodies can be separated from IgG4 and IgG4 Fc using size-exclusion chromatography and non-reducing SDS-PAGE. This method does not rely on a cross-linking immunoassay with its potential for false-positive results due to aggregation and it unambiguously demonstrates that the 'Fab-arm' exchange process depends entirely on the Fc part (hinge + CH2 + CH3). (C) 2010 Elsevier Ltd. All rights reserved.
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