Cost-effective production of recombinant human interleukin 24 by lactose induction and a two-step denaturing and one-step refolding method

被引:10
作者
Amirzada, Muhammad Imran [1 ]
Yu, Minglei [2 ]
Gong, Xiaohai [2 ]
Chen, Yun [2 ]
Zhu, Ruiyu [2 ]
Lei, Jianyong [2 ]
Jin, Jian [2 ,3 ]
机构
[1] Jiangnan Univ, Sch Biotechnol, Key Lab Ind Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China
[2] Jiangnan Univ, Sch Pharmaceut Sci, Wuxi 214122, Jiangsu, Peoples R China
[3] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Hlth Sci, Shanghai, Peoples R China
关键词
rhIL24; Cytokine; Lactose induction; LC-MS/MS; HepG2; HETEROLOGOUS PROTEINS; ESCHERICHIA-COLI; MDA-7/IL-24; GROWTH; OPTIMIZATION;
D O I
10.1007/s10295-013-1367-2
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Recombinant human interleukin 24 (rhIL24) is a member of the interleukin 10 (IL10) family of cytokines with novel therapeutic properties. Human IL24 possesses three N glycosylation sites and a disulfide bridge. The cost and composition of culture media is critical for commercial-scale production of recombinant proteins in E. coli. Addition of yeast extract and glucose to medium enhances rhIL24 production, and the use of lactose instead of IPTG for induction drops the cost and decreases toxicity. In addition, a two-step denaturing and one-step refolding (2DR) strategy improves rhIL24 production. The 2DR strategy replaces a more conventional approach for protein solubilization and refolding. LC-MS/MS provides definitive identification and quantitative information on rhIL24. Single-step purified rhIL24 displayed biological activity on HepG2 hepatocellular carcinoma cells, but no effect on L02 cells. Proliferation analysis suggests that rhIL24 may have potential use as a medication. In the present study, we developed a simple process for producing quality product with high purity. The expression and purification of rhIL24 described here may be a step towards inexpensive large-scale production.
引用
收藏
页码:135 / 142
页数:8
相关论文
共 26 条
[1]   Therapeutic Applications of Interleukin 24 (IL24): A Review [J].
Amirzada, Muhammad Imran ;
Jin, Jian .
TROPICAL JOURNAL OF PHARMACEUTICAL RESEARCH, 2012, 11 (06) :1023-1027
[2]  
[Anonymous], 2002, ASS AD VECT SAF TOX, V13, P3
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   Effect of induction conditions on production and excretion of Aeromonas hydrophila chitinase by recombinant Escherichia coli [J].
Chen, SJ ;
Chang, MC ;
Cheng, CY .
JOURNAL OF FERMENTATION AND BIOENGINEERING, 1997, 84 (06) :610-613
[5]   Production of MBP-HepA fusion protein in recombinant Escherichia coli by optimization of culture medium [J].
Chen, Yin ;
Xing, Xin-Hui ;
Ye, Fengchun ;
Kuang, Ying ;
Luo, Mingfang .
BIOCHEMICAL ENGINEERING JOURNAL, 2007, 34 (02) :114-121
[6]  
Cosat FP, 2011, BRIT J CANCER, V105, P640
[7]  
Dehaghani S.A., 2010, INT J ENV SCI DEV, V1, P111
[8]   Quantitative analysis of acrylamide labeled serum proteins by LC-MS/MS [J].
Faca, Vitor ;
Coram, Marc ;
Phanstiel, Doug ;
Glukhova, Veronika ;
Zhang, Qing ;
Fitzgibbon, Matthew ;
McIntosh, Martin ;
Hanash, Samir .
JOURNAL OF PROTEOME RESEARCH, 2006, 5 (08) :2009-2018
[9]   Structural Mapping of Post-translational Modifications in Human Interleukin-24 ROLE OF N-LINKED GLYCOSYLATION AND DISULFIDE BONDS IN SECRETION AND ACTIVITY [J].
Fuson, Kerry L. ;
Zheng, Mingzhong ;
Craxton, Molly ;
Pataer, Abujiang ;
Ramesh, Rajagopal ;
Chada, Sunil ;
Sutton, R. Bryan .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2009, 284 (44) :30526-30533
[10]   RETRACTED: Strategies for efficient production of heterologous proteins in Escherichia coli (Retracted article. See vol. 98, pg. 5787, 2014) [J].
Jana, S ;
Deb, JK .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2005, 67 (03) :289-298