Skip to main content

Generation of hepatitis B virus PreS2-S antigen in Hansenula polymorpha

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.


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.

This is a preview of subscription content, access via your institution.


  • 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.

    CAS  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • 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.

    Article  CAS  PubMed  Google Scholar 

  • Sjogren M H. 2005. Prevention of hepatitis B in nonresponders to initial hepatitis B virus vaccination. Am J Med, 118Suppl 10A: 34S–39S.

    Article  CAS  PubMed  Google Scholar 

  • Torbenson M, Thomas D L. 2002. Occult hepatitis B. Lancet Infect Dis, 2: 479–486.

    Article  PubMed  Google Scholar 

  • 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)

    Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding authors

Correspondence to Jianqiang Li or Hanmei Xu.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

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).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • hepatitis B virus (HBV)
  • PreS2-S
  • virus-like particle (VLP)
  • Hansenula polymorpha