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Construction of the Antibody Library and Selection of the Antibodies Specifically Binding to the Prostate Carcinoma Cells
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Abstract:
Objective: To obtain phage-displayed ScFv library directly against prostate carcinoma cells, and select antibodies binding to prostate carcinoma cells specifically, so as to lay a foundation for developing diagnostic agents and clinical therapies of prostate carcinoma.Methods:Balb/c mice were immunized i.p .with purified membrane protein mixture of prostate carcinoma cells PC3, DU145. mRNA was isolated from the spleens of immunized mice, heavy and light chain genes （VH and VL） of antibody were amplified separately by RT-PCR and assembled into ScFv gene with a specially constructed linker DNA., the ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated sample was transformed into competent E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage. After five rounds of panning with PC3 cells, the positive clones were selected with the ELISA from the enriched phages. Results: A ScFv library of 3.5×10 6 was obtained and one phage-ScFv which can bind specifically PC3 cells was found. Conclusions: A prostate carcinoma specific antibody was identified ,which paves a way for study of prostate carcinoma.
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Clc Number:
R73-3
