[毕赤酵母表达的重组人白细胞介素11的纯化与鉴定]

[Purification and characterization of recombinant human interleukin 11 which expressed by Pichia pastoris].

作者信息

Huang Y S, Dong Y, Li H, Wang T Y, Qiu J W, Yu Y N

机构信息

School of Medicine, Zhejiang University, Hangzhou 310005, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2001 May;17(3):250-3.

PMID:11517594

Abstract

This study first time report a method to purify the rhIL-11 which expressed by Pichia pastoris. rhIL-11 was secreted into the supernatant and collected by centrifugation. The purity of rhIL-11 reached 97% through the steps of ultrafiltration, SP Sepharose FF, Phenyl Sepharose HP and Sephadex G25. Analysis of SDS-PAGE, Western-blotting, IEF, RP-HPLC, Mass spectrometer, N and C terminus amino acid sequence and bioactivity was conducted. All the analysis results proved that the rhIL-11 expressed by Pichia pastoris was the same as Neumeg which was expressed in E. coli with fusion expression system. So it is possibly a cheaper and easier method to produce rhIL-11 for clinical use.

摘要

本研究首次报道了一种纯化毕赤酵母表达的重组人白细胞介素-11（rhIL-11）的方法。rhIL-11分泌至上清液中，通过离心收集。经超滤、SP Sepharose FF、Phenyl Sepharose HP和Sephadex G25步骤后，rhIL-11的纯度达到97%。进行了SDS-PAGE、Western印迹、IEF、RP-HPLC、质谱仪、N端和C端氨基酸序列及生物活性分析。所有分析结果证明，毕赤酵母表达的rhIL-11与在大肠杆菌中利用融合表达系统表达的Neumeg相同。因此，这可能是一种生产用于临床的rhIL-11的更廉价、更简便的方法。