重组蜂毒素-基因变构IL-2的纯化与生物活性

Purification and Biological Activity of Recombinant Melittin- ^(88)ArgIL-2

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。

Objective To prepare purified recombinant melittin-^88ArglL-2 chimeric protein and analyze its biological activity. Methods Transform the constructed recombinant plasmid pGEX-4T-2/melittin-^88ArglL-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.