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Updated: Jun 6, 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 monoclonal antibodies for human BCSC-1 protein]
Fang Yuan1, Ji-Yu Ju, Yan-Fei Liu
1Department of Immunology, Weifang Medical University; Key Laboratory of Immunology in Universities of Shandong, Weifang 261053, China. yuanfangchina@yahoo.com.cn
Aim:
To obtain monoclonal antibodies against human BCSC-1 protein for further study of the structure and function of human BCSC-1 protein.
Methods:
pET-30a-BCSC-1 plasmid was constructed and transformed into E.coli BL21 (DE3) to express recombinant protein. BALB/c mice were immunized with recombinant human BCSC-1 protein, hybridoma cell lines secreting monoclonal antibodies against human BCSC-1protein were screened by regular cell fusion and subcloning approach. The specificity of these monoclonal antibodies were determined by ELISA and Western blotting. Expression of BCSC-1 protein in pcDNA3.1/v5-HisB-BCSC-1transfected MCF-7 cells was identified by Immunohistochemistry.
Results:
Successfully constructed a prokaryotic expression vector pET30a-BCSC-1.BCSC-1 protein was expressed in E.coli BL21 (DE3). One hybridoma cell line 1B3 stable in secreting specific monoclonal antibodies wa successfully obtained. It could bind specifically to BCSC-1 protein proved by Western blotting and Immunohistochemistry assay.
Conclusion:
Monoclonal antibodies of high specificity against human BCSC-1 protein has been successfully prepared, which laid the foundation for the further study of human BCSC-1 protein.
