【摘 要】
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Human epididymal protease inhibitor (eppin) may be effective as a male contraceptive vaccine.In a number of studies,eppin with an engineered His6-tag has been produced using prokaryotic expression systems.For production of pharmaceutical-grade proteins fo
【机 构】
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School of Radiation Medicine and Public Health, Medical College of Soochow University, Suzhou 215123
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Human epididymal protease inhibitor (eppin) may be effective as a male contraceptive vaccine.In a number of studies,eppin with an engineered His6-tag has been produced using prokaryotic expression systems.For production of pharmaceutical-grade proteins for human use,however,the His6-tag must be removed.This study describes a method for producing recombinant human eppin without a His6-tag.We constructed plasmid pET28a (+)-His6-tobacco etch virus (TEV)-eppin for expression in Escherichia coll.After purification and refolding,the fusion protein His6-TEV-eppin was digested with TEV protease to remove the His6-tag and was further purified by NTA-Ni2+ affinity chromatography.Using this procedure,2 mg of eppin without a His6-tag was isolated from 1 I of culture with a purity of ＞95％.The immunogenicity of the eppin was characterized using male Balb/c mice.
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