title: Soluble expression of recombinant human SMP30 for detecting serum SMP30 antibody levels in hepatocellular carcinoma patients.
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.
authors.affiliation: School of Pre-clinical Sciences, Guangxi Medical University, Nanning, China.
authors.forename: Sheng-Chang
authors.initials: SC
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Huang
authors.forename: Yong-Xiang
authors.initials: YX
authors.lastname: Zhao
authors.forename: Shu-Yan
authors.initials: SY
authors.lastname: Liu
authors.forename: Shu-Jia
authors.initials: SJ
authors.lastname: He
authors.forename: Xiao-Xun
authors.initials: XX
authors.lastname: Xie
authors.forename: Gou-Rong
authors.initials: GR
authors.lastname: Luo
authors.forename: Su-Fang
authors.initials: SF
authors.lastname: Zhou
mesh_terms: D000912:Antibodies, Neoplasm / Q000097:blood*; D015415:Biomarkers / Q000097:blood*; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000097:blood / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000097:blood / Q000276:immunology*; D008113:Liver Neoplasms / Q000097:blood / Q000276:immunology*; D009367:Neoplasm Staging; D010957:Plasmids; D011379:Prognosis; D011994:Recombinant Proteins / Q000097:blood / Q000276:immunology*
chemical_list: D000912:Antibodies, Neoplasm; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; C513323:RGN protein, human; D011994:Recombinant Proteins