Preparation and Characterization of SA-hIL2 Fusion Protein

China Biotechnology

2009, Vol. 29

Issue (07): 12-16 DOI:

Preparation and Characterization of SA-hIL2 Fusion Protein

ZHANG Lin1|HU Zhi-ming2|FA Ping-ping2|LIANG Zhong-kun2|GAO Ji-min1,2

1 Zhejiang Provincial Key Laboratory of Medical Genetics, Wenzhou Medical College, Wenzhou325035, China 2 Institute of Biotherapy, School of Biotechnology, Southern Medical University, Guangzhou510515, China

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Abstract Abstract Objective: To prepare and characterize streptavidin-tagged human interleukin-2 (SA-hIL2) fusion protein. Methods: pET24a-6His-SA-hIL2 plasmid was constructed and expressed in BL 21(DE3) host bacteria to generate fusion protein. The recombinant fusion protein SA-hIL2 was purified through the Ni-NTA affinity chromatography, and then refolded. The efficiency of surface modification of the fusion protein on the biotinylated B16.F10 tumor cells was evaluated by a flow cytometer. CCK-8 method was used to determine the proliferating effect of SA-hIL2 fusion protein on human peripheral-blood lymphocyte (PBL) cells stimulated by PHA. Results: The recombinant SA-hIL2 fusion protein was highly expressed in BL21(DE3) at up to 20% of total bacterial proteins. The fusion protein SA-hIL2 exhibited the bi-functionality: proliferation promoting activity of hIL-2 on PBL cells, and SA-mediated high-affinity binding to the biotinylated surface of B16.F10 cells with about 95% surface modification efficiency. Conclusion: SA-IL2 bi-functional fusion protein was generated, which will make feasible the development of hIL2-surtface-modified cancer cell vaccine.

Key words： Interleukin-2 Streptavidin Fusion protein Surface modification

Received: 14 January 2009 Published: 28 July 2009

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	ZHANG Lin-1
	HU Zhi-Meng-2
	FA Ping-Ping-2
	LIANG Zhong-Gun-2
	GAO Ji-Min-1
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ZHANG Lin-1, HU Zhi-Meng-2, FA Ping-Ping-2, LIANG Zhong-Gun-2, GAO Ji-Min-1, 2. Preparation and Characterization of SA-hIL2 Fusion Protein. China Biotechnology, 2009, 29(07): 12-16.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2009/V29/I07/12

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