Premium
Isolation of HLA and tumor antigens by means of affinity chromatography employing anti‐β 2 ,‐microglobulin (β 2 m) antiserum
Author(s) -
Rauch Joyce E.,
Shuster Joseph,
Thomson David M. P.,
Gold Phil
Publication year - 1978
Publication title -
cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/1097-0142(197809)42:3+<1601::aid-cncr2820420838>3.0.co;2-v
Subject(s) - antiserum , affinity chromatography , human leukocyte antigen , sepharose , antigen , beta 2 microglobulin , radioimmunoassay , immunoprecipitation , antibody , microbiology and biotechnology , chemistry , virology , medicine , biochemistry , biology , immunology , enzyme
A method for the isolation of HLA antigen molecules from normal and cancerous solid human tissue is described. The method employs anti‐β 2 ‐microglobulin (β 2 m) antiserum coupled to Sepharose beads as an immunosorbent affinity medium. The anti‐β 2 m affinity chromatography procedure greatly purifies and selectively enriches HLA and any material that copurifies by affinity, with β 2 m and /or HLA molecules. The HLA isolated by this purification procedure was used to immunize rabbits. The antisera obtained were absorbed on β 2 m to remove all anti‐β 2 m antibody activity. The use of such anti‐HLA antisera in radioimmunoassays, immunoprecipitation studies, and F(ab′) 2 blocking experiments demonstrated that these antisera are directed against a common HLA determinant present on the heavy (alloantigen‐bearing) chain of all HLA molecules. The use of an identical procedure employing human tumor tissues has resulted in the isolation of HLA‐like or HLA‐associated tumor‐specific antigens as demonstrated by the leukocyte adherence inhibition (LAI) assay.