In vitro immunization of rat spleen lymphocytes with Forssman glycosphingolipid antigen and the generation of a monoclonal antibody
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Date
1991-03Author
Gathuru, JK
Miyoshi, I
Naiki, M
Type
ArticleLanguage
enMetadata
Show full item recordAbstract
A procedure for in vitro immunization of splenic lymphocytes with a glycolipid antigen is described. Culture medium supernatant of ConA- and PHA-stimulated spleen cells and that of Con A-stimulated human Jurkat T cell line (IL-2-rich medium) were used as sources of cytokines to support T and B cell stimulation, and anti-mu was used to support B cell differentiation. Unprimed rat spleen cells (2 x 10(6)/ml) were stimulated with 2 micrograms/ml Forssman glycolipid antigen coupled to Sepharose for 4 days. The cells were fused with a mouse myeloma cell line P3-X63-Ag8-U1. At initial screening, 12% of the colony forming wells were secreting specific antibody. After cloning, a stable hybridoma cell line (designated 4C3) was established which secreted a monoclonal IgM antibody directed against the carbohydrate moiety of Forssman glycosphingolipid (GalNAc alpha 1-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc-ceramide).
URI
http://hinari-gw.who.int/whalecomwww.ncbi.nlm.nih.gov/whalecom0/pubmed/2010622http://erepository.uonbi.ac.ke:8080/xmlui/handle/123456789/31802
Citation
J Immunol Methods. 1991 Mar 1;137(1):95-102.Publisher
University of Nairobi School of Public Health, University of Nairobi
Collections
- Faculty of Health Sciences (FHS) [10377]