[Preparation of rhIL-11 from fusion protein by using enterokinase].
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.
Zhao
Yang
Y
National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China.
Huang
He
H
D004765:Enteropeptidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D017370:Interleukin-11 / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D017370:Interleukin-11; D011993:Recombinant Fusion Proteins; D004765:Enteropeptidase