Biosintesis Antigen Permukaan Hepatitis B “HBsAg100” pada Escherichia coli dalam Rangka Produksi Protein Rekombinan sebagai Model Imunogen untuk Menghasilkan Antibodi

Authors

  • Slamet Riyadi Student of Veterinary Science Study Program, School of Postgraduate Studies, Bogor Agricultural University
  • Rarah RA Maheswari Faculty of Animal Husbandry, Bogor Agricultural University
  • Mirnawati Sudarwanto Faculty of Veterinary Science, Bogor Agricultural University
  • Fransiska RZ Fakulty of Technology Agricultural, Bogor Agricultural University
  • Muhamad Ali Chairman Laboratory of Microbiology & Biotechnology Faculty of Animal Husbandry, University of Mataram, Lombok

https://doi.org/10.33476/jky.v18i2.188

Keywords:

Ekspresi protein, protein rekombinan, purifikasi, HBsAg100

Abstract

Biosintesis protein rekombinan melalui Escherichia coli memberikan alternatif untuk menghasilkan protein antigen yang bermanfaan bagi kepentingan kesehatan yang bebas dari protein manusia. Penelitian ini menggabungkan fragmen DNA dari antigen permukaan virus Hepatitis B dengan gen penyandi enzim gluthation-S-transferase (GST) di dalam plasmid p GEX-4T-2 yang di ekspresikan di dalam sel-sel Escherichia coli. Polypeptida dengan berat molekul sekitar 34,8 kDa telah diproduksi dan diidentifikasi sebagai protein gabungan GST-HB100. Protein gabungan tersebut kemudian dimurnikan menggunakan kolum GSTrap yang disambung dengan kolum HiTrap. Selanjutnya, protein hasil pemurnian tersebut diharapkan bisa digunakan sebagai bahan vaksin atau untuk menghasilkan antibodi.

References

Hu WG, Wei J, Yang XX, Xia HC, Li F, and Zhang ZC 2004. Expression of overlapping Pre-S1 Fragment Recombinant Proteins for the determination of immunogenic domains in HBsAg PreS1 region. Acta Biochimica et Biophysica Sinica 36 (6): 397-404.

Joshi N, and Kumar A 2001. Immunoprophylaxis of hepatitis B virus infection. Indian J. Med. Microbiol.19: 172-183.

Joung YH, Youm JW, Jeon JH, Lee BC, Ryu JC, Hong HJ et al., 2004. Expression of the hepatitis B surface S and preS2 antigens in tubulers of Solanum tuberosum. Plant Cell Rep. 22: 925-930.

Kimura T, Ohno N, Terada N, Rokuhara A, Matsumoto A, Yagi S et al., 2005. Hepatitis B virus DNAnegative Dane particles lack core protein

but contain a 22-kDa precore protein without C-terminal arginine-rich domain. J. Biolo. Biochem. 280: 2171321719.

Koschorreck M, Fischer M, Barth S, and Pleiss J 2005. How to find soluble proteins: a comprehensive analysis of alfa/beta hydrolases for recombinant expression in E. coli. BMC Genomics 6: 1-10.

Kristensen J, Petersen HUS, Mortensen KK, Sorensen HP 2005. Generation of monoclonal antibodies for the assesment of protein purification by recombinant ribosomal coupling. Int. J. Biol. Macromolecules 37: 212-217.

Lok ASF, and McMahon BJ 2001. Chronic hepatitis B. Hepatology, 34, 1225-1241.

Lombardi A, Sperandei M, Cantale C, Giacomini P, Galeffi P 2005. Fungtional expression of a singlechain antibody specific or the HER2 human oncogene in a bacterial reducing environment. Protein Expr. Purif. 44: 10-15.

Maeng CY, Oh MS, Park IH, Hong HJ 2001. Purification and structural analysis of the hepatitis B virus preS1 expressed from Escherichia coli. Biochem. Biophys. Res. Com. 282: 787-792.

Mulyanto, Soewignjo S, Gunawan S, Sumarsidi D, Kadir S, and Wiryo H 2002. Hepatitis B seroprevalence among children in Mataram, Indonesia: following a seven-year mass immunization program. Report meeting of the USJapan cooperative medical science program asian region collaboration research project 2001, Shanghai.

Soewignjo S, dan Mulyanto 1984. Epidemiologi infeksi virus Hepatitis B di Indonesia. Acta Medica Indonesiana 15: 215-230.

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Published

2010-07-27

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Section

Research Articles