Immunochemical studies on a panosyl-azoprotein conjugate

Abstract

The trisaccharide panose, 6-O-[alpha]-d-glucopyranosyl-4-O-[alpha]-d-glucopyranosyl-d-glucose, coupled to bovine serum albumin (BSA) via an azophenyl linkage was used as an artificial antigen to immunize rabbits. Antisera rendered specific for the introduced haptenic grouping by absorption with whole calf serum were studied by immunodiffusion, quantitative precipitation and hapten inhibition. Anti-hapten showed a specificity directed against the introduced p-phenylazo-[ss]-panoside group. Hapten inhibition assays employing various di- and triglucosides, including two structural isomers of panose, established the relative importance of an [alpha]-configuration and a nonreducing terminal (1,6)-(1,4) ordered sequence of glucoside bonds for optimal interaction with antibody. A heterogeneity in the specificity of antibodies formed in response to [ss]-panosyl-BSA was evidenced from cross reactions obtained between anti-panoside and various sugar azoprotein conjugates.