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DETECTION EFFICACY OF SOLUBLE HLA‐A, B ANTIGENS USING 1D‐IEF
Author(s) -
Kubens B. S.,
Päbler M.,
GrosseWilde H.
Publication year - 1994
Publication title -
international journal of immunogenetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.41
H-Index - 47
eISSN - 1744-313X
pISSN - 1744-3121
DOI - 10.1111/j.1744-313x.1994.tb00220.x
Subject(s) - isoelectric focusing , human leukocyte antigen , antigen , monoclonal antibody , serology , immunoprecipitation , proband , antibody , microbiology and biotechnology , biology , hla a , immunology , chemistry , gene , biochemistry , mutation , enzyme
SUMMARY Soluble HLA class I alloantigens (sHLA class I) can be typed according to their isoelectric points (IEP) after immunoprecipitation by w6/32 monoclonal antibody (mAb) coupled to immunomagnetic beads and focusing. In order to prove the large scale efficacy of this methodology, EDTA‐plasma samples from 344 probands HLA‐A, B typed by serology were analysed by one‐dimensional isoelectric focusing and HLA class I specific Westernblot (1D‐IEF). In addition, detergent solubilized HLA class I membrane molecules from approximately one half of the probands were studied too. Soluble HLA‐A24, B7, B18, B62 antigens were identified in nearly all experiments, whereas A28, B13, and B51 could be detected in about 50%. A third group of HLA antigens (A26, B8, B44) could be visualized rarely. The difficulties of detection might be due to the different affinity of mAb w6/32 to certain sHLA class I gene products or to variable amounts of sHLA class I in the plasma specimens. Some modifications of the antigen capture technique have already led to a slightly better degree of antigen recognition in 25 probands tested. Thus, HLA‐A, B typing using sHLA molecules and 1D‐IEF in the assay format presented does not yet seem to be a definitive alternative for HLA class I serology or biochemistry of membrane‐bound HLA class I molecules but it should be a promising technique if no cells are available or donor‐derived sHLA allotypes are to be monitored after HLA mismatched organ transplantation.