title: C1q binding by a high affinity anti-fluorescein murine monoclonal IgM antibody and monomeric subunits.
abstract: Comparative interactions of purified rabbit C1q with 18-2-3, a high affinity (2-3 X 10(10) M-1) anti-fluorescein (anti-F1) murine monoclonal IgM antibody (pentamer) and constitutive monomeric subunits (IgMs) were studied. Using a solid phase radioimmunoassay (SPRIA), based on immobilized polyvalent antigen, it was shown that the mechanism of C1q binding to IgM was characteristically multiphasic while IgMs yielded monophasic binding curves. The latter compared qualitatively and quantitatively with a monoclonal IgG2a anti-fluorescein antibody with the same intrinsic affinity of 2-3 X 10(10) M-1. C1q binding efficiency to antibodies was significantly enhanced when the immunoglobulins interacted with immobilized multivalent antigen. Monoclonal IgM antibody bind identically to six F1-carrier protein conjugates independent of epitope (F1) density. In contrast, the C1q-antibody interaction binding was dependent upon epitope density. An average distance between F1 epitopes of 80 A was optimal for C1q binding by IgM. At low concn of IgM, when fluorescein was bound by antigen-binding sites on adjacent subunits of an intact pentamer, C1q appeared to bind IgM intramolecularly.
authors: Department of Microbiology, University of Illinois, Urbana 61801.
authors: S M
authors: SM
authors: Swanson
authors: M A
authors: MA
authors: Dombrink-Kurtzman
authors: E W
authors: EW
authors: Voss
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003166:Complement Activating Enzymes / Q000302:isolation & purification; D003172:Complement C1 / Q000302:isolation & purification; D015922:Complement C1q; D000939:Epitopes / Q000032:analysis; D019793:Fluorescein; D005452:Fluoresceins / Q000276:immunology; D007075:Immunoglobulin M / Q000145:classification; D011863:Radioimmunoassay
chemical_list: D000911:Antibodies, Monoclonal; D003172:Complement C1; D000939:Epitopes; D005452:Fluoresceins; D007075:Immunoglobulin M; D015922:Complement C1q; D003166:Complement Activating Enzymes; D019793:Fluorescein