摘要
为了获得高活性高纯度的rh IFNα2b,对重组人干扰素α2b进行克隆、表达,并深入研究了其纯化工艺。采用重叠延伸PCR法合成了编码IFNα2b的基因,用DNA重组技术构建了原核表达载体p BV220-IFNα2b,获得了稳定的工程菌种。发酵产物通过破菌、洗涤获得包涵体,再经过变性、复性、离子交换层析和凝胶过滤层析的纯化,得到rh IFNα2b纯品,其比活可达1×10^8IU/mg。实验结果为进一步开展临床前研究和长效制剂奠定了基础。
Recombinant human interferon α2b was prepared by cloning,expression,purification for obtaining rh IFN α2b with high activity and purity. Overlap extension PCR was performed to obtain the gene encoding for IFN α2b. A prokaryotic vector expressing rh IFN α2b was constructed successfully. The expression vector p BV220-IFN α2b was transformed into E. coli strain DH5α. After fermentation,the bacteria cells was collected and lysed. Finally,the rh IFN α2b was purified by denaturation,renaturation,ion exchange chromatography and gel filtration chromatography,and its specific activity reached 1×10^8IU / mg. The results was expected to lay the foundation for preclinical research and long-acting preparation.
出处
《生物技术进展》
2016年第3期212-218,共7页
Current Biotechnology
关键词
干扰素Α2B
制备
表达
纯品
interferon α2b
preparation
expressing
purify
