Immunological characterization of xenogenic anti-metatype antibodies.
High affinity murine anti-fluorescein monoclonal antibody (Mab) 4-4-20 (K alpha = 1.3 x 10(10) M-1; IgG2a; kappa), affinity labeled with fluorescein isothiocyanate (I), served as the immunogen to elicit a xenogenic (rabbit) polyclonal "anti-metatype" reagent in a rabbit specific for the liganded state. The reagent did not bind Mab 4-4-20 in its nonliganded (idiotypic) conformation and demonstrated no reactivity with fluorescein ligand. Interaction of the anti-metatype reagent with liganded 4-4-20 caused a significant decrease in the dissociation rate of fluorescyl ligand bound to Mab 4-4-20 and a single-chain antibody derivative of 4-4-20. Additionally, most members of the 4-4-20 idiotype family displayed the same kinetic effect in the presence of anti-metatype. Members most closely related to Mab 4-4-20 (based on concentration in Mab required to inhibit the 4-4-20 idiotype/4-4-20 anti-idiotype interaction 50%) showed relatively greater decreases in the dissociation rate, establishing a quantitative correlation between idiotype and metatype. The immunologically distinct metatypic state is discussed in terms of conformational changes in or near the active site upon binding ligand, possibly involving two amino acid residues which "close off" the mouth of the antibody-active site upon binding fluorescein.
Weidner
K M
KM
Department of Microbiology, University of Illinois, Urbana 61801.
Voss
E W
EW
D000345:Affinity Labels; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000910:Antibodies, Heterophile / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015478:Antigens, Heterophile / Q000276:immunology; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013861:Thiocyanates
D000345:Affinity Labels; D000888:Antibodies, Anti-Idiotypic; D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D015478:Antigens, Heterophile; D005452:Fluoresceins; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate