Preference for IgG mAb binding insulin in solution or on surfaces is related to immunoglobulin variable region structures.

Tikhomirov OY, Thomas JW
J Autoimmun. 1997 10 (6): 541-9

PMID: 9451593 · DOI:10.1006/jaut.1997.0161

Studies on insulin autoantibodies often show a lack of correlation between enzyme-linked immunosorbent assays (ELISAs) and fluid phase radioimmunoassays (RIAs). Similarly, a set of IgG anti-insulin monoclonal antibodies (mAb) from BALB/c mice are found to differ in their binding in ELISAs and RIAs. To understand the structural basis for differential insulin binding, soluble and complexed biotinylated insulin is used to confirm binding properties independent of insulin-coated plastic and radioiodination. The binding properties of intact mAb are also present in Fab fragments, indicating ligand preference is not related to valence or to the Fc portion of Ig. Analysis of binding to soluble or bound ligand in relationship to antibody variable (V) region structures indicates that differential binding in the two assays is a property of heavy chain variable region structure. Studies also show that limited amino acid replacements arising during maturation of the immune response may change the binding preference for an individual mAb. These findings indicate that differences in detection of insulin binding in solid phase and fluid phase are not artefactual but reflect intrinsic structural features of immunoglobulin interaction with insulin.

MeSH Terms (20)

Amino Acid Sequence Animals Antibodies, Monoclonal Biotin Enzyme-Linked Immunosorbent Assay Female Humans Immunoglobulin Fragments Immunoglobulin G Immunoglobulin Variable Region Insulin Mice Mice, Inbred BALB C Molecular Sequence Data Phenotype Radioimmunoassay Sequence Homology, Amino Acid Solutions Structure-Activity Relationship Surface Properties

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