Abstract: Recombinant human interferon α2b was prepared by cloning, expression, purification for obtaining rhIFN α2b with high activity and purity. Overlap extension PCR was performed to obtain the gene encoding for IFN α2b. A prokaryotic vector expressing rhIFN α2b was constructed successfully. The expression vector pBV220-IFN α2b was transformed into E. coli strain DH5α. After fermentation, the bacteria cells was collected and lysed. Finally, the rhIFN α2b was purified by denaturation, renaturation, ion exchange chromatography and gel filtration chromatography, and its specific activity reached 1×108 IU/mg. The results was expected to lay the foundation for preclinical research and long-acting preparation.

Key words: interferon α2b, preparation, expressing, purify

摘要： 为了获得高活性高纯度的rhIFN α2b，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IFN α2b的基因，用DNA重组技术构建了原核表达载体pBV220-IFN α2b，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rhIFN α2b纯品，其比活可达1×108 IU/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。

关键词: 干扰素&alpha, 2b, 制备, 表达, 纯品