Conformational isomerism of IgG antibodies.
The purpose of this study was to determine why apparently homogeneous IgG antibodies were, in some cases, fractionated into at least two components by liquid-liquid partition chromatography (LLPC) in an aqueous two-phase system. Four mouse monoclonal IgG antibodies, two against albumin, one against IgG and one against thyroxine, were shown to adopt different conformational isomeric forms. The four antibodies existed in an equilibrium between two or three conformational forms, the proportion of which could also be estimated by LLPC. Since LLPC detects mainly conformational differences within the antigen-binding sites of IgG antibodies, it could be concluded that the conformational forms differed with respect to their combining sites. Moreover, the isomeric forms of an antibody directed against a protein antigen, formed antigen-antibody complexes with almost identical surface properties. In contrast, complexes with different surface properties were formed when the hapten or hapten conjugated to BSA was bound. Thus, both the conformational isomers could bind antigen, at least when the antigen was a small hapten or a hapten conjugated to a carrier protein. Our results suggest that six out of 57 monoclonal IgG antibodies exist in equilibrium between at least two conformational forms and the biological significance of this isomerism is discussed.
Hansson
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UB
Department of Biochemistry, Lund University, Sweden. Ulla-Britt.Hansson@biokem.lu.se
Wingren
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Alkner
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D001666:Binding Sites, Antibody; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D007525:Isoelectric Focusing; D051379:Mice; D011487:Protein Conformation; D012709:Serum Albumin / Q000276:immunology; D012710:Serum Albumin, Bovine / Q000276:immunology; D013974:Thyroxine / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D013974:Thyroxine