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GUO Wei, WANG Chen, WANG Yuheng, GAO Xiangdong. Expression, purification and functional assay of rhIL23R-CHR[J]. Journal of China Pharmaceutical University, 2013, 44(6): 583-588. DOI: 10.11665/j.issn.1000-5048.20130619
Citation: GUO Wei, WANG Chen, WANG Yuheng, GAO Xiangdong. Expression, purification and functional assay of rhIL23R-CHR[J]. Journal of China Pharmaceutical University, 2013, 44(6): 583-588. DOI: 10.11665/j.issn.1000-5048.20130619
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Expression, purification and functional assay of rhIL23R-CHR

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  • The IL23R-CHR gene was cloned from human leukocyte cDNA library and inserted into pMD18T vector. The recombinant plasmid was confirmed by DNA sequencing. Followed, the IL23R-CHR was subcloned into pET22b/pET28a/pET32a vector, and then the recombinant expression plasmids were transformed into BL21(DE3). In this study, we optimized the expression condition by comparing direct expression between soluble fusion protein expression, secretion expression and inclusion body expression. Finally, hIL23R-CHR was expressed and purified from Trx-hIL23R-CHR fusion protein with a purity of 90%. Affinity assay also demonstrated that purified hIL23R-CHR was able to bind to human IL-23. Histological examination revealed that hIL23R-CHR can protect the central nervous system of EAE mice.
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