[Cloning, expression and bio-activity assay of chimeric fusion protein sTNFRII-IgG Fc].
Tumor necrosis factor (TNF) is a key cytokine in immunology system and is related to many human diseases. In order to inhibit the activity of TNF, cDNA coding for soluble TNF receptor II (sTNFRII) and human IgG Fc were linked using a flexible hinge. This gene was expressed in E. coli as a chimeric protein and purified by metal chelate chromatography. The results show that the fusion protein exists in the physiological form as a dimer, has the ability to bind with TNF and inhibits the cytotoxicity of TNF on L929 cells. Contrasting to monomer sTNFRII, the chimeric protein has an improved bioactivity, and displays potential prospects for research and application.
Xu
Chun-Xiao
CX
State Key Laboratory of Molecular Virology and Genetic Engineering, Institute of Virology, Beijing 100052, China.
Yao
Li-Hong
LH
Zu
Dong
D
Chen
Ai-Jun
AJ
Huang
Guo-Jin
GJ
Zhang
Zhi-Qing
ZQ
D015703:Antigens, CD / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western / Q000379:methods; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D015870:Gene Expression; D005818:Genetic Engineering; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007074:Immunoglobulin G / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011995:Recombination, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D015703:Antigens, CD; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D018124:Receptors, Tumor Necrosis Factor; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha