Production of oligosaccharide-binding monoclonal antibodies of diverse specificities by immunization with purified tumor-associated glycolipids inserted into liposomes with lipid A
✍ Scribed by Thomas Brodin; Jan Thurin; Niklas Strömberg; Karl-Anders Karlsson; Hans Olov Sjögren
- Publisher
- John Wiley and Sons
- Year
- 1986
- Tongue
- English
- Weight
- 650 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0014-2980
No coin nor oath required. For personal study only.
✦ Synopsis
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(Pl+ 3)Gal( a1 --f 3)Gal( f3l+ 3)Glc(P1+ l)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.