Abstract
Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen (HBsAg) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus (HBV) PreS2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsAg, particularly in terms of cytotoxic T lymphocyte (CTL) reaction. In the current study, the HBV PreS2-S gene encoding an extra 26 amino acids (PreS2 C-terminus) located at the N-terminus of HBsAg was cloned into the pVCH1300 expression vector. PreS2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purification steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The final product was obtained with a total yield of ∼15% and purity of ∼99%. In keeping with previous studies, ∼22 nm viruslike particles were detected using electron microscopy. The generated PreS2-S antigen will be further studied for efficacy and safty in animals.
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References
Awan A R, Zahoor M Y, Javed M M, Babar M E, Saleem Z. 2012. Expression of pres2/S antigen of hepatitis B virus isolated from Pakistan in yeast cells. Pak. J. Bot., 44: 355–359.
Cabral G A, Marciano-Cabral F, Funk G A, Sanchez Y, Hollinger F B, Melnick J L, Dreesman G R. 1978. Cellular and humoral immunity in guinea pigs to two major polypeptides derived from hepatitis B surface antigen. J Gen Virol, 38: 339–350.
Chen H, Chuai X, Deng Y, Wen B, Wang W, Xiong S, Ruan L, Tan W. 2012. Optimisation of prime-boost immunization in mice using novel protein-based and recombinant vaccinia (Tiantan)-based HBV vaccine. PLoS One, 7: e43730.
Chi S W, Kim D H, Kim J S, Lee M K, Han K H. 2006. Solution conformation of an immunodominant epitope in the hepatitis B virus preS2 surface antigen. Antiviral Res, 72: 207–215.
Faber K N, Haima P, Harder W, Veenhuis M, Ab G. 1994. Highly-efficient electrotransformation of the yeast Hansenula polymorpha. Curr Genet, 25: 305–310.
G. Birkenmeyer L. 2003. Hepatitis B virus: life cycle and morphogenesis. In: Ias K M (ed.), Perspectives in Medical Virology, vol. Volume 10. Elsevier, p109–125.
Gellissen G, Janowicz Z A, Weydemann U, Melber K, Strasser A W, Hollenberg C P. 1992. High-level expression of foreign genes in Hansenula polymorpha. Biotechnol Adv, 10: 179–189.
Hadiji-Abbes N, Borchani-Chabchoub I, Triki H, Ellouz R, Gargouri A, Mokdad-Gargouri R. 2009. Expression of HBsAg and preS2-S protein in different yeast based system: a comparative analysis. Protein Expr Purif, 66: 131–137.
Hardy E, Martinez E, Diago D, Diaz R, Gonzalez D, Herrera L. 2000. Large-scale production of recombinant hepatitis B surface antigen from Pichia pastoris. J Biotechnol, 77: 157–167.
Huang Y, Bi J, Zhang Y, Zhou W, Li Y, Zhao L, Su Z. 2007. A highly efficient integrated chromatographic procedure for the purification of recombinant hepatitis B surface antigen from Hansenula polymorpha. Protein Expr Purif, 56: 301–310.
Janssen H L, van Zonneveld M, Senturk H, Zeuzem S, Akarca U S, Cakaloglu Y, Simon C, So T M, Gerken G, de Man R A, Niesters H G, Zondervan P, Hansen B, Schalm S W. 2005. Pegylated interferon alfa-2b alone or in combination with lamivudine for HBeAg-positive chronic hepatitis B: a randomised trial. Lancet, 365: 123–129.
Kuroda S, Fujisawa Y, Iino S, Akahane Y, Suzuki H. 1991. Induction of protection level of anti-pre-S2 antibodies in humans immunized with a novel hepatitis B vaccine consisting of M (pre-S2 + S) protein particles (a third generation vaccine). Vaccine, 9: 163–169.
Langley K E, Egan K M, Barendt J M, Parker C G, Bitter G A. 1988. Characterization of purified hepatitis B surface antigen containing pre-S(2) epitopes expressed in Saccharomyces cerevisiae. Gene, 67: 229–245.
Lian M, Zhou X, Chen B, Li C, Gu X, Luo M, Zheng X. 2008. Identification of the critical regions in hepatitis B virus preS required for its stability. J Pept Sci, 14: 307–312.
Liaw Y F, Leung N W, Chang T T, Guan R, Tai D I, Ng K Y, Chien R N, Dent J, Roman L, Edmundson S, Lai C L. 2000. Effects of extended lamivudine therapy in Asian patients with chronic hepatitis B. Asia Hepatitis Lamivudine Study Group. Gastroenterology, 119: 172–180.
Milich D R, Thornton G B, Neurath A R, Kent S B, Michel M L, Tiollais P, Chisari F V. 1985. Enhanced immunogenicity of the pre-S region of hepatitis B surface antigen. Science, 228: 1195–1199.
Niederau C, Heintges T, Lange S, Goldmann G, Niederau C M, Mohr L, Haussinger D. 1996. Long-term follow-up of HBeAg-positive patients treated with interferon alfa for chronic hepatitis B. N Engl J Med, 334: 1422–1427.
Sjogren M H. 2005. Prevention of hepatitis B in nonresponders to initial hepatitis B virus vaccination. Am J Med, 118Suppl 10A: 34S–39S.
Torbenson M, Thomas D L. 2002. Occult hepatitis B. Lancet Infect Dis, 2: 479–486.
Jiang Z G, Wang Y, Shao X A, Yue Y, Hong X W, Xu L, Xiong S D. 2007. Antibodized gene vaccine harboring to HBV PreS2/S gene can enhance HBV specific CTL response induced by gene immunization. Chinese J Immunol, 963–967. (In Chinese)
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Xu, X., Ren, S., Chen, X. et al. Generation of hepatitis B virus PreS2-S antigen in Hansenula polymorpha . Virol. Sin. 29, 403–409 (2014). https://doi.org/10.1007/s12250-014-3508-9
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DOI: https://doi.org/10.1007/s12250-014-3508-9
Keywords
- hepatitis B virus (HBV)
- PreS2-S
- virus-like particle (VLP)
- Hansenula polymorpha