重组蜂毒素-基因变构IL-2的纯化与生物活性

目的制备纯化重组蜂毒素-基因变构IL-2(Melittin-88ArgIL-2)嵌合蛋白,并检测其生物活性。方法将含表达载体的大肠杆菌DH5α,经IPTG诱导表达。表达产物经离子交换层析与亲和层析进行纯化,SDS-PAGE鉴定,MTT染色法检测嵌合蛋白对Hela细胞增殖的抑制作用。结果所表达的可溶性蛋白,纯化后纯度达95%,蛋白浓度0.5g/L。纯化后的嵌合蛋白在体外能抑制Hela细胞的增殖。结论已成功地制备并纯化了重组melittin-88ArgIL-2嵌合蛋白,该蛋白具有生物活性,为中试放大和纯化工艺研究奠定了基础。

Purification and Biological Activity of Recombinant Melittin-88 ArgIL-2

Objective To prepare purified recombinant melittin- 88ArgIL-2 chimeric protein and analyze its biological activity.Methods Transform the constructed recombinant plasmid pGEX-4T-2/melittin- 88ArgIL-2 to E.coli DH5α for expression under induction of IPTG.Purify the expressed product by ion exchange and affinity chromatography and identify by SDS-PAGE.Determine the inhibitory effect of expressed chimeric protein on the proliferation of Hela cells by MTT method.Results The purity and protein concentration of expressed soluble protein after purification reached 95% and 0.5 g/L respectively.The purified chimeric protein,at a concentration of 20 μg/L,showed significant inhibitory effect in vitro on the proliferation of Hela cells.Conclusion The purified recombinant melittin- 88ArgIL-2 chimeric protein was successfully prepared and showed biological activity.It laid a foundation of scale-up of pilot procedure and development of purification procedure of the chimeric protein.