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Two-dimensional gel electrophoresis of material immunoprecipitated from radiolabeled cells by anti-HLA class II monoclonal antibodies has been a useful technique but the patterns observed have varied considerably between laboratories. In this study we investigated the parameters that vary between different laboratories. We found that heterogeneity due to post synthetic modification of beta and alpha chains of DR and DQ can be avoided by labeling cells for 30 min with 35S-methionine. A mixture of ampholytes pH range 3.5-10:5-7 of 1:4 run for 3000 V/h gave the best resolution of beta and alpha chains. With this technique we could clearly reveal differences in the DR beta-2 chains of those DR2 haplotypes, Dw2, Dw12 and LD'tb24. The Dr beta 1 position varied between different DR types.

Type

Journal article

Journal

Tissue Antigens

Publication Date

08/1986

Volume

28

Pages

72 - 83

Keywords

Antibodies, Monoclonal, Antibody Specificity, Cell Line, Electrophoresis, Polyacrylamide Gel, Histocompatibility Antigens Class II, Humans, Isoelectric Point, Macromolecular Substances, Molecular Weight, Polymorphism, Genetic