Expression and purification of hepatitis B surface antigen S from Escherichia coli; A new simple method

被引:9
作者
Elghanam M.S. [1 ]
Attia A.S. [1 ]
Shoeb H.A. [1 ]
Hashem A.E.M. [1 ]
机构
[1] Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo
关键词
GST-fusion; HBsAg; Hepatitis B; Purification;
D O I
10.1186/1756-0500-5-125
中图分类号
学科分类号
摘要
Abstract. Background: Hepatitis B is a liver disease primarily caused by hepatitis B virus (HBV) infection. It is distributed worldwide and associated with high mortality and morbidity rates. HBV infections can be avoided by the administration of the currently available vaccine and can be easily diagnosed through commercially available kits. Both the vaccine and the diagnostic kits depend on using the hepatitis B surface antigen (HBsAg) as an antigen. Developing countries such as, Egypt, suffer from the widespread of HBV infections and the limited resources to provide adequate supplies of either the vaccine or the diagnostic kits. Therefore the need for an easy, rapid, low cost method to produce HBsAg is urgently needed within this setting. Findings. To achieve this goal, the gene encoding the HBsAg(S) protein was cloned and expressed as a fusion protein with a GST tag in Escherichia coli. The recombinant protein was successfully expressed and purified in both good quality and quantity. Conclusions: The simplified and the relatively low cost of the used protocol make this an attractive alternative to protocols currently used for the purification of HBsAg(S). The exploiting of this achievement for new diagnostics can be directed for application in the developing countries where they are extremely needed. © 2012 Attia et al; licensee BioMed Central Ltd.
引用
收藏
相关论文
共 26 条
[1]  
Hou J., Liu Z., Gu F., Epidemiology and prevention of hepatitis B virus infection, International Journal of Medical Sciences, 2, 1, pp. 50-57, (2005)
[2]  
Lavanchy D., Hepatitis B virus epidemiology, disease burden, treatment, arid current and emerging prevention and control measures, Journal of Viral Hepatitis, 11, 2, pp. 97-107, (2004)
[3]  
Neurath A.R., Kent S.B.H., Strick N., Hepatitis B virus contains pre-S gene-encoded domains, Nature, 315, 6015, pp. 154-156, (1985)
[4]  
Alberti A., Pontisso P., Milanesi G., Methods for the study of pre-S proteins of hepatitis B virus and their antibodies: Pathogenetic and clinical implications, La Ricerca in Clinica e in Laboratorio, 18, pp. 241-258, (1988)
[5]  
Bo H., Minjian L., Guoqiang H., Zhaoxia L., Zhenyir Z., Lin L., Expression of hepatitis B virus S gene in Pichia pastoris and application of the product for detection of anti-HBs antibody, Journal of Biochemistry and Molecular Biology, 38, 6, pp. 683-689, (2005)
[6]  
Walsh K., Alexander G.J.M., Update on chronic viral hepatitis, Postgraduate Medical Journal, 77, 910, pp. 498-505, (2001)
[7]  
Kobayashi M., Asano T., Utsunomiya M., Itoh Y., Fujisawa Y., Nishimura O., Kato K., Kakinuma A., Recombinant hepatitis B virus surface antigen carrying the pre-S2 region derived from yeast:purification and characterization, J Biotechnol, 8, pp. 1-22, (1988)
[8]  
El-Sherif A.M., Abou-Shady M.A., Al-Hiatmy M.A., Al-Bahrawy A.M., Motawea E.A., Screening for hepatitis B virus infection in Egyptian blood donors negative for hepatitis B surface antigen, Hepatol Int, 1, pp. 469-470, (2007)
[9]  
El-Zayadi A.-R., Ibrahim E.H., Badran H.M., Saeid A., Moneib N.A., Shemis M.A., Abdel-Sattar R.M., Ahmady A.M., El-Nakeeb A., Anti-HBc screening in Egyptian blood donors reduces the risk of hepatitis B virus transmission, Transfusion Medicine, 18, 1, pp. 55-61, (2008)
[10]  
Watson N., A new revision of the sequence of plasmid pBR322, Gene, 70, pp. 399-403, (1988)