Purification of anti-Lec antibodies with specificity for beta DGal(1 replaced by 3)beta DGlcNAcO- using a synthetic...

Artificial antigens: Antibodies were raised in rabbits with beta DGal(1 linked to 3)beta DGlcNAc-BSA and purified by absorption elution on a column of the synthetic oligosaccharide covalently bound to a silicate instead of BSA. The purified antibodies agglutinated specifically erythrocytes from Le (a-b-) nonsecretor donors (Lec). Natural antigens: Antibodies were raised in goats using boiled saliva from O/O, le/le, se/se donors. The population of antibodies which was absorbed on a column consisting of beta DGal(1 linked to 3)etaDGlcNAc-O-R (type 1 chain precursor disaccharide) groupings attached to a silicate was collected and then refined by passage through a column which consisted of similarly immobilized alpha LFuc(1 linked to 2)beta DGal(1 linked to 3)betaGDlcNAc-(H type 1), beta(DGal(1 linked to 3)-[alpha LFuc(1 linked to 4)]beta DGlcNAc-(Lea), and beta DGal(1 replaced by 4)beta DGlcNAc-(type 2 core chain precursor) groupings. The refined antibodies proved to have a reaction pattern with salivas that is the same as that previously reported for anti-Lec sera. The data obtained with natural and artificial antigens are thus compatible with the biosynthetic pathway for the formation of the Lewis antigenic determinants proposed by Graham and suggest that the Lec antigen may simply be the type 1 precursor chain which is required for the formation of the Lea, Led and Leb antigenic determinants.