Production of oligosaccharide-binding monoclonal antibodies of diverse specificities by immunization with purified tumor-associated glycolipids inserted into liposomes with lipid A.
Two species of neutral glycosphingolipids purified from rat colon carcinoma tissue, isoglobotetraosylceramide [GalNAc(beta 1----3)Gal(alpha 1----3)Gal(beta 1----3)Glc(beta 1----1)Cer] and a related 6-sugar "analogue" were inserted into liposomes together with lipid A (from bacterial lipopolysaccharide) and used for immunization of mice and monoclonal antibody production. The yield of hybridomas producing glycolipid-specific antibody was 5-10% using a high-dose booster schedule with liposome-inserted glycolipid. In contrast the frequency was below 0.1% (no glycolipid-binding antibodies were found) when using the previously described method of immunizing with glycolipid coated on the surface of acid-treated S. minnesota. Monoclonal antibodies were screened on the purified glycolipids used for immunization and selected for differential reactivity to the two glycolipids. A diversity of specificities was demonstrated by binding to the purified antigens, in a thin-layer chromatogram binding assay and in binding tests to tumor and normal target cells.
D000230:Adenocarcinoma / Q000032:analysis / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000951:Antigens, Neoplasm; D003110:Colonic Neoplasms / Q000032:analysis / Q000276:immunology; D006017:Glycolipids / Q000276:immunology*; D008050:Lipid A / Q000276:immunology*; D008081:Liposomes; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011920:Rats, Inbred WF
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D006017:Glycolipids; D008050:Lipid A; D008081:Liposomes