Antigenic studies on an enzymatically sialylated carbohydrate: NeuAc(alpha 2-3)Gal(beta 1-3)GalNAc.
Sialic acid residues are often the end moiety of the carbohydrate chain of biologically important glycoconjugates. It is difficult to study sialylated glycoconjugates because the purification of these compounds is often laborious yielding only very small amounts of oligosaccharides for study. Chemical synthesis of sialylated compounds is complicated by the labile nature of the sialic acid bond. In both of these cases the sialylated compounds would need to be conjugated to a polypeptide to be an effective immunogen, and again, such conjugation is fraught with problems due to the instability of the sialic acid linkage. We have developed a combined enzymatic and synthetic route for obtaining quantities of sialylated carbohydrates conjugated to a protein carrier in amounts sufficient for antigenic studies. The notable novelty of this protocol is the addition of sialic acid after the carbohydrate-protein conjugation step. Antiserum to the compounds was developed and after absorption, antibodies that demonstrate a requirement for sialic acid for their binding were produced and studied. CA 125 has been shown to be a prognostically significant marker for ovarian adenocarcinoma. The nature of the epitope involved has been analyzed with conflicting results. To attempt to resolve this conflict, we initiated studies on sialylated antigens with NeuAc alpha 2-3Gal beta 1-3GalNAc. This trisaccharide occupies the terminal region in a series of complex carbohydrates which have been suggested to be involved as the epitope. Hanisch et al. reported that the neuraminic acid was important for the reaction.
Henion
T
T
Department of Gynecologic Oncology, Roswell Park Cancer Institute, Buffalo, New York 14263.
Handley
D
D
Vargas
F
F
Matta
K L
KL
Piver
M S
MS
Diakun
K R
KR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology / Q000378:metabolism; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D006801:Humans; D008969:Molecular Sequence Data; D009438:Neuraminic Acids / Q000378:metabolism; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000378:metabolism; D012799:Sialyltransferases / Q000276:immunology / Q000378:metabolism*; D014312:Trisaccharides / Q000138:chemical synthesis* / Q000378:metabolism; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015295:Antigens, Tumor-Associated, Carbohydrate; D009438:Neuraminic Acids; C064068:Thomsen-Friedenreich antigen, cryptic; D014312:Trisaccharides; D012710:Serum Albumin, Bovine; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase