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| Soluble Expression of Human Leukemia Inhibitory Factor in Prokaryotic Cells and Its Purification |
| ZHANG He-ming1,2, CAI Chu-fan1,2, LIU Yang1,2, GAN Long-zhan1,2, JIAO Xue-miao3, TIAN Yong-qiang1,2 |
1. Department of Biomaterials and Leather Engineering, College of Textiles and Food Science, Sichuan University, Chengdu 610065, China;
2. Key laboratory of Leather Chemistry and engineering(Sichuan University), Ministry of Education, Chengdu 610065, China;
3. Key Laboratory of Plant Stress Biology, Henan University, Kaifeng 475001, China |
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Abstract Numerous studies have demonstrated that leukemia inhibitory factor (LIF) is one of the most important polyfunctional cytokine, hLIF is a pleiotropic cytokine with multiple effects on different types of cells and tissues, its unique biological characteristics make it widely used. The homemade hLIF which is biologically active is viesented, hLIF gene was cloned into pET32a, and it successfully expressed the soluble recombinant protein in E. coli using the thioredoxin (Trx) protein as a fusion partner. After purification based on membrane adsorber technology, the fusion protein was cleaved using EK protease. Released, soluble hLIF was subsequently purified by cation exchange, The samples analysis by SDS-PAGE and Western blot. This procedure yields up to 4.75mg rhLIF with 98.1% purity. Functional analysis of the purified rhLIF by murine myeloblastic leukemia M1 cell proliferation assay demonstrates biological activity that is similar and comparable to that of hLIF, and with the EC50 of 6ng/ml and the corresponding specific activity of 0.5×107 IU/mg.
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Received: 11 April 2017
Published: 25 September 2017
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