Determination of affinity constants and the number of binding sites of monoclonal antibodies specific for southern bean mosaic virus

Abstract

Values of the equilibrium dissociation constant and the number of attachment sites for the binding of radiolabeled F AB fragments prepared from four monoclonal antibodies to southern bean mosaic virus have been determined. Monoclonal antibodies B6 and B10 produced against the bean type strain of southern bean mosaic virus (SBMV-B) and 1C4 and 1D6 produced against the cowpea type strain (SBMV-C) were more reactive with the untreated virus than with EDTA-swollen particles. B6 and B10 reacted with 180 and 90 sites, respectively, on the SBMV-B particle and were not reactive with SBMV-C. Antibody 1D6 reacted with 180 sites on both SBMV-B and SBMV-C, whereas 1C4 bound to 180 sites on SBMV-C and 60 sites on SBMV-B. The equilibrium dissociation constants for B10, 1C4, and 1D6 were similar but antibody B6 bound with an 8- to 10-fold lower affinity. Solid phase competitive antibody inhibition analysis showed that 1C4 and 1D6 were mutually inhibitory and that these antibodies also inhibited the binding of B6 and B10. Antibody B6 partially inhibited the binding of 1D6 but did not affect 1C4 or B10 binding. No inhibition of B6, 1C4, or 1D6 binding was observed when B10 was the competing antibody. Possible sites of reaction of these monoclonal antibodies on the SBMV particle are discussed.