Immunochemical studies on a laminaribiosyl-azoprotein conjugate

Abstract

Abstraet-Laminaribiose (3-o-/3-D-glucopyranosyl-D-glucose) was coupled to bovine serum albumin (BSA) by an azophenyl linkage to provide the synthetic antigen BSAp-phenylazo-/3-1aminaribioside. The behavior of antisera prepared in rabbits immunized with the /3-1aminaribiosyl conjugate was examined by immunodiffusion, quantitative precipitation and hapten inhibition. Anticonjugate absorbed with carrier protein showed the greatest reactivity with the homologous /3-1aminaribiosyl-BSA antigen, but also showed some cross precipitation with /3-cellobiosyl, /3-sophorosyl and /3-gentiobiosyl-BSA conjugates. Glucobioses linked through the /3-(1-* 2), /3-(1-* 3), /3-(1-* 4) and /3-(1-* 6) positions, as well as laminaridextrins and tri and tetrasaccharides of/3-1inked glucose possessing a laminaribiose moiety either at a nonreducing end location or at a subterminal location, were assayed for their ability to inhibit antilaminaribioside precipitation. Hapten inhibition data showed anticonjugate to possess a high degree of specificity directed against the terminal nonreducing/3-1aminaribiosyl end group.

Immunochemical studies on a panosyl-azoprotein conjugate

“…Melibiose, 6-O-a-D-galactopyranosyl-D-glucose, which differs from isomaltose in the configuration of the hydroxyl at C-4' of the non-reducing end, has only one thirty-sixth the molar inhibitory ability of its C-4' epimer, isomaltose. This finding is comparable to similar effects obtained with glucose, galactose, maltose, gentiobiose, lactose, sophorose and laminaribiose azoproteinconjugates (Avery and Goebel, 1929;Goebel et al, 1934;Allen et al, 1967Allen et al, , 1970Karush, 1957).…”

“…When the panosyl-antipanosyl system was examined by immunodiffusion, multiple banding was observed with certain concentrations of reactants. A recent study by Henney (1970) may have some relevance to this finding and could explain similar observations reported with other sugar azoprotein systems (Allen et al, 1970;Gleich and Allen, 1965). Sera of guinea pigs immunized with DNP-human gammaglobulin show the presence of antibodies whose specificity includes both hapten and part of the carrier termed 'hapten-carrier complex specific', as well as antibodies specifi~ f~,c tLe h,ptcn itself (Henney, 1970).…”

“…In addition to sharing the maltosyl unit as a common structural feature, fl-maltosyl and /3-panosyl BSA conjugates also share the phenylazo group and may even possess identical phenylazo peptides. The contribution to immunochemical reactivity of the aglycone structure of phenylazo-protein-sugar conjugates has been well established for several other systems (Beiser et al, 1960;Allen et al, 1967Allen et al, , 1970Gleich and Alien, 1965;Tanenbaum et al, 1961;Karush, 1957) and is similarly involved in the cross reactivity found with/3-maltosyl BSA.…”

“…Immunochemical studies on sugar-protein conjugates involving the relationship of sugar hapten structure to antihapten specificity have been carried out almost entirely with mono and disaccharides as haptens (Avery and Goebel, 1929;Goebel et al, 1934;Goebel, 1939;Beiser et al, 1960;Arakatsu et al, 1966;Allen et al, 1967;Martineau et al, 1969;Allen et al, 1970). Apart from a study by Arakatsu et al (1966), employing a synthetic antigen consisting of isomaltotrionic acid coupled to bovine serum albumin (BSA), conjugates possessing oligosaccharide determinants made up of three or more sugar residues have received little attention.…”

Quantitative studies on the interaction of concanavalin A, the carbohydrate-binding protein of the jack bean, with model carbohydrate-protein conjugates

“…They were able to show mono-and oligosaccharides are immunogenic when coupled protein and that the antibodies produced exhibited a specificity towards the introduced sugar. Since then a large number of carbohydrate-protein conjugates have been prepared (Allen et al, 1970;Arbkatsu et al, 1966;Gieich and Allen, 1965;Goldstein et al, 1967;Karush, 1957;Beiser et al, 1960).…”