[Prokaryotic expression of endostatin and preparation of polyclonal antibody].

BACKGROUND & OBJECTIVE: The aim of this study was to express and purify human endostatin and to prepare polyclonal antibody of mouse anti-human endostatin. METHODS: The cDNA of endostatin was amplified by PCR, then recombined into prokaryotic expression vector and transformed into Escherichia coli BL21 for expression; the mice were immunized with purified products. RESULTS: Prokaryotic expression vector pQE-30 of human endostatin was successfully constructed; the expression product was gained after pQE-30 was transferred into BL21. After purified by Ni affinity chromatography, the product was identified to be a single component by SDS-PAGE. Western blot analysis showed that high titer mouse anti-human endostatin polyclonal antibody was successfully prepared. CONCLUSION: Highly purified expression product and prepared polyclonal antibody provide the necessary material for further study.