Browse > Article
http://dx.doi.org/10.7314/APJCP.2013.14.4.2383

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)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.14, no.4, 2013 , pp. 2383-2386 More about this Journal
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.
Keywords
SMP30; genetic engineering; soluble expression; molecular chaperone;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Baneyx F, Mujacic M (2004). Recombinant protein folding and misfolding in Escherichia coli. Nat Biotechnol, 22, 1399-408.   DOI   ScienceOn
2 de Marco A, Deuerling E, Mogk A, Tomoyasu T, Bukau B (2007) . Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli. BMC Biotechnol, 7, 32.   DOI
3 Diamant S, Eliahu N, Rosenthal D, Goloubinoff P (2001). Chemical chaperones regulate molecular chaperones in vitro and in cells under combined salt and heat stresses. J Biol Chem, 276, 39586-91.   DOI   ScienceOn
4 Frank B, Jennifer M, Petersen R (2010). A cell-based screen for inhibitors of protein folding and degradation. Cell Stress Chaperon, 15, 913-27.   DOI
5 Goloubinoff P, Gatenby AA, Lorimer GH (1989). GroE heatshock proteins promote assembly of foreign prokaryotic ribulose bisphosphate carboxylase oligomers in Escherichia coli. Nature, 337, 44-7.   DOI   ScienceOn
6 Hoffmann F, Rinas U (2004). Roles of heat-shock chaperones in the production of recombinant proteins in Escherichia coli. Adv Biochem Eng Biotechnol, 89,143-61.
7 Huang L, Shakhnovich EI (2012). Is there an en route folding intermediate for Cold shock proteins? Protein Sci, 21, 677-85.   DOI
8 Kiefhaber T, Rudolph R, Kohler HH, Buchner J (1991). Protein aggregation in vitro and in vivo: a quantitative model of the kinetic competition between folding and aggregation. Biotechnology, 9, 825-9.   DOI   ScienceOn
9 Moonsuk SC, Saxena AM, Chilukuri NW (2010). A strategy for the production of soluble human senescence marker protein-30 in Escherichia coli. Biochem Bioph Res Co, 393, 509-13.   DOI   ScienceOn
10 Nishihara K, Kanemori M, Yanagi H (2000). Overexpression of trigger factor prevents aggregation of recombinant proteins in Escherichia coli. Appl Environ Microbiol, 66, 884-9.   DOI   ScienceOn
11 Oyunsuren T, Sanduijav R, Davaadorj D, Nansalmaa D (2006). Hepatocellular carcinoma and its early detection by AFP testing in Mongolia. Asian Pac J Cancer Prev, 7, 460-2.
12 Ran F, Gadura N, Michels CA (2010). Hsp90 co-chaperone Aha1 is a negative regulator of the saccharomyces MAL Activator and Acts early in the chaperone activation pathway. J Biol Chem, 285, 13850-62.   DOI   ScienceOn
13 Sambrook J, Russell DW (2001). Molecular cloning: a laboratory manual. Cold Spring Harbor, New York.
14 Schrodel A, de Marco A (2005). Characterization of the aggregates formed during recombinant protein expression in bacteria. BMC Biochem, 31, 10.
15 Timasheff SN (2002). Protein hydration, thermodynamic binding, and preferential hydration. Biochemistry, 41, 13473-82.   DOI   ScienceOn
16 Ventura S, Villaverde A (2006). Protein quality in bacterial inclusion bodies. Trends Biotechnol, 24, 179-85.   DOI   ScienceOn
17 Wilkinson DL, Harrison RG (1991). Predicting the solubility of recombinant proteins in Escherichia coli. Biotechnology, 9, 443-8.   DOI
18 Yamaguchi M, Mori S, Kato S (1988). Calcium-binding protein regucalcin is an activator of ($Ca^{2+}-Mg^{2+}$)-adenosine triphosphatase in the plasma membranes of rat liver. Chem Pharm Bull, 36, 3532-9.   DOI   ScienceOn
19 Zhou SF, Mo FR, Bin YH, et al (2011). Serum immunoreactivity of SMP30 and its tissues expression in hepatocellular carcinoma. Clin Biochem, 44, 331-6.   DOI   ScienceOn
20 Zhou SF, Xie XX, Bin YH (2006). Identification of HCC-22-5 tumor-associated antigen and antibody response in patients. Clinica Chimima Acta, 366, 274-80.   DOI   ScienceOn