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A one-step monoclonal antibody typing procedure that simplifies HLA class I and class II typing
Authors:Jar-How Lee  Mamie Lias  Chun-Tsan Deng  Soudabeh Etessami  Jimmy Loon  Mike Chen  Leninh Banh  Nathan Conger  Dan Connors  Alex Manzo  George Ayoub  Paul I. Terasaki
Affiliation:One Lambda, Inc., Canoga Park, CA, U.S.A.;Department of Surgery, University of California, Los Angeles, CA, U.S.A.
Abstract:
Abstract: We have developed monoclonal antibodies to most HLA specificities, making it possible for us to devise a simple, rapid, one-step micro-cytotoxicity test. The test is performed by adding 1 μl of cells to 1 μl of antibody-complement mixture predotted on the microtest tray. The reactions are read following a 1-hour incubation period (30 minutes in some instances). The analysis of reactions seen on testing 105 class I antibodies and 50 class II antibodies is shown. A comparison of typing by the standard NIH method and the new one-step procedure showed a > 96% concordance in the 500 T cells and 200 B cells we examined. Class I and class II typing could be performed using B cells, thus obviating the need to isolate both T and B cells for HLA typing.
Keywords:MHC    HLA typing    monoclonal antibody    complement    class I    class II    one-step typing
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