Production and characterization of murine monoclonal antibody to human alpha-lactalbumin.
Human alpha-lactalbumin (alpha-LA), a milk protein normally produced by, and restricted to, functionally differentiated breast epithelial cells has been shown immunohistochemically to be a good marker for breast cancer. Antibodies with specificity only for human alpha-LA have not previously been reported. The present study documents the production and characterization of an IgG1 murine monoclonal antibody with specificity restricted only to human alpha-LA. This monoclonal antibody, designated ET-1, was purified by affinity chromatography using human alpha-LA. Specificity of ET-1 for human alpha-LA was established by enzyme linked immunosorbent assay (ELISA), absorption studies, immunoprecipitation and by immunoblotting. ET-1 binds to human alpha-LA with a KD of 7.41 x 10(-8) mol/l and does not cross-react with bovine alpha-LA. This unique reactivity of ET-1, which is not inherently shared by polyclonal antisera, should enable the antibody to be used for the development of a sensitive immunoassay for circulating human alpha-LA.
Thean
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ET
Department of Pathology and Immunology, Monash Medical School, Prahran, Vic., Australia.
Toh
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BH
D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000097:blood; D001943:Breast Neoplasms / Q000097:blood; D005260:Female; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G; D007768:Lactalbumin / Q000097:blood / Q000276:immunology*; D051379:Mice
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D007074:Immunoglobulin G; D007768:Lactalbumin