A method for the preparation and identification of fumitremorgin B-hemisuccinatecarrier protein is described. The overall yield of the fumitremorgin B-hemisuccinate (FTBS) after final purification was 78.6%. The FTBS ...A method for the preparation and identification of fumitremorgin B-hemisuccinatecarrier protein is described. The overall yield of the fumitremorgin B-hemisuccinate (FTBS) after final purification was 78.6%. The FTBS was characterized by UV, IR, EIMS,element analysis, and 1H, 13C NMR. IR was also used to determine the formation of complete antigen complexes. The reaction route was analysized.展开更多
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 pro...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 coli. 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.展开更多
Coated capillary columns were prepared by sol-gel technology and used in the separation of basic proteins with capillary zone electrophoresis. The results indicated that a significant decrease in protein adsorption wa...Coated capillary columns were prepared by sol-gel technology and used in the separation of basic proteins with capillary zone electrophoresis. The results indicated that a significant decrease in protein adsorption was obtained and EOF was also diminished to zero in the pH range of 3-10.展开更多
文摘A method for the preparation and identification of fumitremorgin B-hemisuccinatecarrier protein is described. The overall yield of the fumitremorgin B-hemisuccinate (FTBS) after final purification was 78.6%. The FTBS was characterized by UV, IR, EIMS,element analysis, and 1H, 13C NMR. IR was also used to determine the formation of complete antigen complexes. The reaction route was analysized.
基金ACKNOWLEDGEMENTS The authors are grateful to Dr Zi-Chun Hua （The State Key Laboratory of Pharmaceutical Biotechnology and Department of Biochemistry, College of Life Science, Nanjing University, China） for kindly providing plasmid pET28a （＋） and TEV endoprotease and to Dr Michael G. O＇Rand （Laboratories for Reproductive Biology and Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, USA） for English-language editing. This study was supported by grants from the Key Project of the National Natural Science Foundation of China （No. 30930079） and the Chinese National Nature Science Foundation （No. 81001257）.
文摘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 coli. 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.
文摘Coated capillary columns were prepared by sol-gel technology and used in the separation of basic proteins with capillary zone electrophoresis. The results indicated that a significant decrease in protein adsorption was obtained and EOF was also diminished to zero in the pH range of 3-10.
