Preparation of rhIL-11 from fusion protein by using enterokinase.
- Author:
Yang ZHAO
1
;
He HUANG
Author Information
1. National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Enteropeptidase;
chemistry;
genetics;
metabolism;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Humans;
Interleukin-11;
biosynthesis;
genetics;
Protein Engineering;
methods;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
isolation & purification
- From:
Journal of Experimental Hematology
2008;16(4):903-909
- CountryChina
- Language:Chinese
-
Abstract:
The study was aimed to investigate the technological parameters of rhIL-11 preparation from fusion protein by using enterokinase. Fusion expression vector pET32a/IL-11 was transferred into E.coli BL21 (DE3) pLysS and its expression was induced by IPTG, the lysis supernatant of the engineering strain was purified by Ni-NTA resin and then the target rhIL-11 was digested by auto-cleavaged DsbA-EKL-(His)(8). The results showed that after affinity chromatography, Trx-IL-11 was obtained with the yield of 11.25mg/g, the purity of 89.2% and the recovery of 91.8%. Finally the target rhIL-11 was digested and purified to over 95%. In conclusion, the preparation method of rhIL-11 from fusion protein by using enterokinase is simple and feasible with good separation, which can meet industrial requirements.
