PRODUCTION OF MONOCLONAL-ANTIBODIES TO BRUTONS TYROSINE KINASE

Bruton's X-linked agammaglobulinemia is caused by mutations in a cytop lasmic protein tyrosine kinase termed Bruton's tyrosine kinase (BTK). The protein is expressed in all members of the B cell lineage and is c ritical for B cell development, The protein consists of several module s, including a pleckstrin homology domain and the Src homology domains SH1, SH2, and SH3. We report here the production of monoclonal antibo dies against the pleckstrin homology domain of human BTK. The antibody was produced by immunizing mice with a FLAG-BTK fusion protein, Hybri doma supernatants were screened by ELISA using a GST-BTK fusion protei n as the antigen. Selected monoclonal antibodies recognize denatured B TK on Western blots of peripheral blood mononuclear cell lysates. Mous e BTK protein is also detected. These antibodies should be useful in a ssessing patients with immune deficiency, as well as in studying norma l B cell development.