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Updated: May 30, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Preparation and characterization of anti-human B7-H4 monoclonal antibodies]
Hui Xu1, Peng Fang, Yan-ping Liu
1Biotechnology Institute of Soochow University, Suzhou 215007, China.
Aim:
To prepare anti-B7-H4 monoclonal antibodies (mAbs) and to characterize their biological functions.
Methods:
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.
Results:
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.
Conclusion:
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.
