Unknown

Dataset Information

0

Preparation and immunogenicity of tag-free recombinant human eppin.


ABSTRACT: Human epididymal protease inhibitor (eppin) may be effective as a male contraceptive vaccine. In a number of studies, eppin with an engineered His(6)-tag has been produced using prokaryotic expression systems. For production of pharmaceutical-grade proteins for human use, however, the His(6)-tag must be removed. This study describes a method for producing recombinant human eppin without a His(6)-tag. We constructed plasmid pET28a (+)-His(6)-tobacco etch virus (TEV)-eppin for expression in Escherichia coli. After purification and refolding, the fusion protein His(6)-TEV-eppin was digested with TEV protease to remove the His(6)-tag and was further purified by NTA-Ni(2+) affinity chromatography. Using this procedure, 2 mg of eppin without a His(6)-tag was isolated from 1 l of culture with a purity of >95%. The immunogenicity of the eppin was characterized using male Balb/c mice.

SUBMITTER: Zhang J 

PROVIDER: S-EPMC3739548 | biostudies-other | 2011 Nov

REPOSITORIES: biostudies-other

Similar Datasets

| S-EPMC7416841 | biostudies-literature
| S-EPMC8310097 | biostudies-literature
| S-EPMC7810742 | biostudies-literature
2022-02-16 | GSE196649 | GEO
| S-EPMC10719321 | biostudies-literature
| S-EPMC4016594 | biostudies-literature
| S-EPMC103727 | biostudies-literature
| S-EPMC4185020 | biostudies-literature
| S-EPMC9141723 | biostudies-literature
| S-EPMC7349816 | biostudies-literature