Production of oligosaccharide‐binding monoclonal antibodies of diverse specificities by immunization with purified tumor‐associated glycolipids inserted into liposomes with lipid A

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Abstract
Two species of neutral glycosphingolipids purified from rat colon carcinoma tissue, isoglobotetraosylceramide [GalNAc(β1→3)Gal(α1→3)Gal(β1→3)Glc(β1→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.