ABSTRACT: To prepare anti-B7-H4 monoclonal antibodies (mAbs) and to characterize their biological functions.
A human B7-H4 transfectant cell line L929/B7-H4 was used as an immunogen to immunize BALB/c mice. By means of the B lymphoma hybridoma technique, immunofluorescent cytometry, repeated screening and multiple subcloning, the hybridoma cell lines specifically secreting anti-B7-H4 mAbs were screened. The fast-strip analysis was used to investigate murine Ig subclass. The specificity of mAbs was determined by Dot-blot and Western blot. Competitive inhibition test was employed to identify mAb binding sites on B7-H4. T cell proliferation inhibition blocking test was used to examine mAb biological function.
Two hybridoma cell lines were obtained and named 1F10 and 2B2, respectively. They could secret continuously and stably specific anti-B7-H4 mAbs. The result of Dot-blot indicated that two mAbs could recognize specifically B7-H4, but only mAb 2B2 recognized B7-H4 when using Western blot. The competitive inhibition test showed that two mAbs bound different epitopes on B7-H4. The two mAbs could partially block the inhibitory effects of B7-H4 on T cell proliferation in vitro.
Two hybridoma cell lines secreting anti-B7-H4 mAbs were obtained. Obtained two mAbs provided useful tool for further studying B7-H4's biological functions.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 08/2011; 27(8):887-90.