Preparation and immunogenicity of tag-free recombinant human eppin

• 来源期刊: 亚洲男性学杂志（英文版）
• 年，卷（期）: 2011,（6）
• 页码范围: 889-894
• 页数: 6页
• 语种: 英文
• DOI: 10.1038/aja.2011.89

摘要

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.