Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Soluble Expression of Recombinant Human Smp30 for Detecting Serum Smp30 Antibody Levels in Hepatocellular Carcinoma Patients

  • Zhang, Sheng-Chang (School of Pre-clinical Sciences, Guangxi Medical University) ;
  • Huang, Peng (Central Laboratory of Genetic and Metabolism, MCH Hospital of Guangxi Autonomous Region) ;
  • Zhao, Yong-Xiang (Biological Targeted Diagnosis and Treatment Center of Guangxi Medical University) ;
  • Liu, Shu-Yan (School of Pre-clinical Sciences, Guangxi Medical University) ;
  • He, Shu-Jia (School of Pre-clinical Sciences, Guangxi Medical University) ;
  • Xie, Xiao-Xun (School of Pre-clinical Sciences, Guangxi Medical University) ;
  • Luo, Gou-Rong (School of Pre-clinical Sciences, Guangxi Medical University) ;
  • Zhou, Su-Fang (School of Pre-clinical Sciences, Guangxi Medical University)
  • Published : 2013.04.30
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Abstract

Senescence marker protein 30 (SMP30), a hepatocellular carcinoma (HCC) associated antigen, was earlier shown by our research group to be highly expressed in HCC paracancerous tissues, but have low levels in HCC tissues. In order to detect anti-SMP30 antibody in serum of HCC patients, we established pET30a-SMP30 and pColdIII-SMP30 expression systems in Escherichia coli. However, the expression product was mainly in the form of inclusion bodies. In this research, we used several combinations of chaperones, four molecular chaperone plasmids with pET30a-SMP30 and five molecular chaperone plasmids with pColdIII-SMP30 to increase the amount of soluble protein. Results showed that co-expression of HIS-SMP30 with pTf16, combined with the addition of osmosis-regulator, and a two-step expression resulted in the highest enhancement of solubility. A total of 175 cases of HCC serum were studied by ELISA to detect anti-SMP30 antibody with recombinant SMP30 protein. Some 22 were positive and x2 two-sided tests all showed P>0.05, although it remained unclear whether there was a relationship between positive cases and clinical diagnostic data.

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