Monoclonal antibody specific to alpha-2-->3-linked deaminated neuraminyl beta-galactosyl sequence

Fusion of spleen cells from a BALB/c mouse immunized with KDN alpha 2-->3Gal beta 1-->4Glc beta 1-->1Cer ((KDN)GM3) with P3-X63 Ag8.U1 (P3U1) mouse myeloma cells yielded a hybrid cell line that produced monoclonal antibody that bound to (KDN)GM3, but not to Neu5Ac alpha 2-->3Gal beta 1-->4Glc-beta 1-->1Cer ((Neu5Ac)GM3). The specificity of the monoclonal antibody was determined chiefly by the enzyme-linked immunosorbent assay procedure. This antibody was found to react most strongly with (KDN)GM3 and less strongly with a glycoprotein containing a number of KDN alpha 2-->3Gal beta 1-->3-GalNAc alpha 1-->3[8KDN alpha 2-->)n-->6]GalNAc alpha 1-->chains (< n > av = approximately 3). The results indicated that the monoclonal antibody (designated mAb.kdn3G) specifically and effectively recognized a disaccharide structure, KDN alpha 2-->3Gal beta 1-->, and specifically discriminated (KDN)GM3 from (Neu5Ac)GM3. The mAb.kdn3G was used to localize (KDN)GM3 in rainbow trout sperm by the indirect immunofluorescence procedure and the antigen was shown to be mostly, if not completely, associated with the external surface of the entire plasma membrane of rainbow trout sperm. The potential utility of mAb.kdn3G is addressed in searching for KDN-glycoconjugates which contain glycan units having the KDN alpha 2-->3Gal beta 1-->epitope structure.