Recombination of heavy and light chains of immunoglobulins. Use of specific variable and constant region probes to monitor reassociation reactions.
The reassociation of antibody (IgG)-derived heavy (gamma) and light (L) chains was studied by utilizing two specifically localized fluorescent probes. L chains labeled in either the variable domain with fluorescein (specific antigen) or the constant domain with N-(iodoacetamidoethyl)-5-naphthylamine-1-sulfonic acid were prepared. Reassociation of these chains with isolated, unlabeled, gamma chains was monitored by following an increase in the fluorescence polarization of fluorophores at 4 degrees C. Results obtained indicate that the reassociation reaction could be resolved into two distinct steps; the primary interaction of the reactants occurring rapidly, followed by a slower secondary phase possibly representing conformational changes in the variable domains of the reactants after reassociation.
D005452:Fluoresceins; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007142:Immunoglobulin gamma-Chains; D007460:Iodoacetamide / Q000031:analogs & derivatives; D007700:Kinetics; D046911:Macromolecular Substances; D009282:Naphthalenesulfonates; D013050:Spectrometry, Fluorescence
D005452:Fluoresceins; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007142:Immunoglobulin gamma-Chains; D046911:Macromolecular Substances; D009282:Naphthalenesulfonates; D007460:Iodoacetamide