[Preparation and characterization of monoclonal antibodies for human BCSC-1 protein].
OBJECTIVE
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.


CONCLUSIONS
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.
Department of Immunology, Weifang Medical University; Key Laboratory of Immunology in Universities of Shandong, Weifang 261053, China. yuanfangchina@yahoo.com.cn
Fang
F
Yuan
Ji-Yu
JY
Ju
Yan-Fei
YF
Liu
Li-Na
LN
Wang
Da-Lin
DL
DI
Shu-Juan
SJ
Liang
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D045744:Cell Line, Tumor; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D010957:Plasmids / Q000235:genetics
D000911:Antibodies, Monoclonal; D009363:Neoplasm Proteins; C109969:VWA5A protein, human