Establishment of hIL-6 Protein Binding to sIL-6R Model for Screening IL-6 Inhibitors

China Biotechnology

2009, Vol. 29

Issue (11): 60-65 DOI:

Establishment of hIL-6 Protein Binding to sIL-6R Model for Screening IL-6 Inhibitors

SHI Ji-jing¹,LIU Zhao-qi¹,ZOU Kun²,YANG Zu-wei²,GAO Ming-xing²,YANG Fan¹

1.Institute of Molecular Biology, Three Gorges University, Yichang 443002, China
2.Hubei Provincial Key Laboratory of Natural Product Research and Development， Three Gorges University, Yichang 443002, China

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Abstract

AIM: To eatablish the model of hIL-6 protein binding to sIL-6R for screening IL-6 inhibitors. METHODS: Human IL-6 gene was cloned into pET28a(+) prokaryotic expressing vector. The recombinant protein IL-6 was induced by IPTG in E. coli BL21(DE3) and the expressed protein was detected by Western blot assay and ELISA; Human sIL-6R gene was cloned into pET15b prokaryotic expressing vector. The recombinant protein sIL-6R was induced by IPTG in E. coli BL21(DE3) and the expressed protein was detected by Western blot assay.Then IL-6/sIL-6R binding assay was established using enzyme-linked immunosorbent assay (ELISA), and optimized the model through changing the concentrations of IL-6, sIL-6R and IL-6 antibody. RESULTS: Human IL-6 and sIL-6R could be expressed in E. coli BL21(DE3) with high efficiency. The recombinant proteins were characterized by western blot and ELISA.Through optimizing the model,the best conditions were obtained: IL-6R 1?g/well, IL-6 500ng/well, IL-6 antibody 1?g/well.And some components detected by the model had significantly inhibitory effect on IL-6/sIL-6R. CONCLUSION: The screening model of hIL-6 protein binding to sIL-6R was successfully established,which provided a reliable platform for high throughput screening of IL-6 inhibitors.

Key words： IL-6 sIL-6R protein expression ELISA drug screening

Received: 07 April 2009 Published: 07 December 2009

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	SHI Ji-Jing
	LIU Chao-Ai
	JU Kun
	YANG Jie-Wei
	GAO Meng-Xing
	YANG Fan

Cite this article:

SHI Ji-Jing, LIU Chao-Ai, JU Kun, YANG Jie-Wei, GAO Meng-Xing, YANG Fan. Establishment of hIL-6 Protein Binding to sIL-6R Model for Screening IL-6 Inhibitors. China Biotechnology, 2009, 29(11): 60-65.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2009/V29/I11/60


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