Volume 21 Issue 6
Dec.  2008
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QIAN WEN, LI MA, WEI LUO, MING-QIAN ZHOU, XIAO-NING WANG. Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF[J]. Biomedical and Environmental Sciences, 2008, 21(6): 509-513.
Citation: QIAN WEN, LI MA, WEI LUO, MING-QIAN ZHOU, XIAO-NING WANG. Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF[J]. Biomedical and Environmental Sciences, 2008, 21(6): 509-513.
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Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF

  • Institute of Molecular Immunology,Southern Medical University,Guangzhou 510515,Guangdong,China

  • School of Biosciences and Bioengineering,South China University of Technology,Guangzhou 510641,Guangdong,China

Funds:  国家自然科学基金(30771952)%广东省自然科学基金(07117783)%NSFC and the Research Grants Council of Hong Kong Joint Research Scheme(30418003)
  • To study the activities of interleukin (IL)-2 and granulocyte-macrophage colony-stimulating factor (GM-CSF) (hIL-2/mGM-CSF).Methods SOE PCR was used to change the linker of the fusion protein for higher activities.The fusion protein was expressed in Escherichia coli (E.coli) BL21 (DE3) in inclusion body (IB) form.After IB was extracted and clarified,it was denatured and purified by affinity chromatography.The protein was refolded by dilution in a L-arginine refolding buffer and refined by anion chromatography.The protein activity was detected by cytokine-dependent cell proliferation assay Results The expression of hlL-2/mGM-CSF in E.coil yielded approximately 20 mg protein/L culture and the purity was about 90%.The specific activities of IL-2 and GM-CSF were 5.4×106 IU/mg and 7.1×106 IU/mg,respectively.Conclusion This research provides important information about the anti-tumor activity of hIL-2/mGM-CSF in vivo,thus facilitating future clinical research on hIL-2/mGM-CSF used in immune therapy.
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Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF

  • Institute of Molecular Immunology,Southern Medical University,Guangzhou 510515,Guangdong,China
  • School of Biosciences and Bioengineering,South China University of Technology,Guangzhou 510641,Guangdong,China
Funds:  国家自然科学基金(30771952)%广东省自然科学基金(07117783)%NSFC and the Research Grants Council of Hong Kong Joint Research Scheme(30418003)

Abstract: To study the activities of interleukin (IL)-2 and granulocyte-macrophage colony-stimulating factor (GM-CSF) (hIL-2/mGM-CSF).Methods SOE PCR was used to change the linker of the fusion protein for higher activities.The fusion protein was expressed in Escherichia coli (E.coli) BL21 (DE3) in inclusion body (IB) form.After IB was extracted and clarified,it was denatured and purified by affinity chromatography.The protein was refolded by dilution in a L-arginine refolding buffer and refined by anion chromatography.The protein activity was detected by cytokine-dependent cell proliferation assay Results The expression of hlL-2/mGM-CSF in E.coil yielded approximately 20 mg protein/L culture and the purity was about 90%.The specific activities of IL-2 and GM-CSF were 5.4×106 IU/mg and 7.1×106 IU/mg,respectively.Conclusion This research provides important information about the anti-tumor activity of hIL-2/mGM-CSF in vivo,thus facilitating future clinical research on hIL-2/mGM-CSF used in immune therapy.

QIAN WEN, LI MA, WEI LUO, MING-QIAN ZHOU, XIAO-NING WANG. Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF[J]. Biomedical and Environmental Sciences, 2008, 21(6): 509-513.
Citation: QIAN WEN, LI MA, WEI LUO, MING-QIAN ZHOU, XIAO-NING WANG. Expression,Purification,and Refolding of Recombinant Fusion Protein Hil-2/Mgm-CSF[J]. Biomedical and Environmental Sciences, 2008, 21(6): 509-513.
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