1887

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Volume 95, Issue 10

Production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique Free

Abstract

Dengue virus (DENV; genus ) contains a positive-stranded RNA genome. Binding of DENV to host cells is mediated through domain III of the viral envelope protein. Many therapeutic mAbs against domain III have been generated and characterized because of its high antigenicity. We have previously established a novel PCR method named the linear array epitope (LAE) technique for producing monoclone-like polyclonal antibodies. To prove this method could be utilized to produce antibody against epitopes with low antigenicity, a region of 10 aa (VNIEAEPPFG) from domain III of the envelope protein in DENV serotype 2 (DENV2) was selected to design the primers for the LAE technique. A DNA fragment encoding 10 directed repeats of these 10 aa for producing the tandem-repeated peptides was obtained and fused with glutathione -transferase (GST)-containing vector. This fusion protein (GST-Den EIII-His) was purified from and used as antigen for immunizing rabbits to obtain the polyclonal antibody. Furthermore, the EIII antibody could recognize envelope proteins either ectopically overexpressed or synthesized by DENV2 infection using Western blot and immunofluorescence assays. Most importantly, this antibody was also able to detect DENV2 virions by ELISA, and could block viral entry into BHK-21 cells as shown by immunofluorescence and quantitative real-time PCR assays. Taken together, the LAE technique could be applied successfully for the production of antibodies against antigens with low antigenicity, and shows high potential to produce antibodies with good quality for academic research, diagnosis and even therapeutic applications in the future.

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© 2014 The Authors
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2014-10-01
2024-04-25
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References

  1. Barratt-Boyes S. M., Vlad A., Finn O. J. 1999; Immunization of chimpanzees with tumor antigen MUC1 mucin tandem repeat peptide elicits both helper and cytotoxic T-cell responses. Clin Cancer Res 5:1918–1924[PubMed]
     [Google Scholar]
  2. Beasley D. W., Barrett A. D. 2002; Identification of neutralizing epitopes within structural domain III of the West Nile virus envelope protein. J Virol 76:13097–13100 [View Article][PubMed]
     [Google Scholar]
  3. Bhardwaj S., Holbrook M., Shope R. E., Barrett A. D., Watowich S. J. 2001; Biophysical characterization and vector-specific antagonist activity of domain III of the tick-borne flavivirus envelope protein. J Virol 75:4002–4007 [View Article][PubMed]
     [Google Scholar]
  4. Cecilia D., Gould E. A. 1991; Nucleotide changes responsible for loss of neuroinvasiveness in Japanese encephalitis virus neutralization-resistant mutants. Virology 181:70–77 [View Article][PubMed]
     [Google Scholar]
  5. Chávez J. H., Silva J. R., Amarilla A. A., Moraes Figueiredo L. T. 2010; Domain III peptides from flavivirus envelope protein are useful antigens for serologic diagnosis and targets for immunization. Biologicals 38:613–618 [View Article][PubMed]
     [Google Scholar]
  6. Chiu M. W., Yang Y. L. 2003; Blocking the dengue virus 2 infections on BHK-21 cells with purified recombinant dengue virus 2 E protein expressed in Escherichia coli. Biochem Biophys Res Commun 309:672–678 [View Article][PubMed]
     [Google Scholar]
  7. Chokephaibulkit K., Perng G. C. 2013; Challenges for the formulation of a universal vaccine against dengue. Exp Biol Med (Maywood) 238:566–578 [View Article][PubMed]
     [Google Scholar]
  8. Crill W. D., Chang G. J. 2004; Localization and characterization of flavivirus envelope glycoprotein cross-reactive epitopes. J Virol 78:13975–13986 [View Article][PubMed]
     [Google Scholar]
  9. Crill W. D., Roehrig J. T. 2001; Monoclonal antibodies that bind to domain III of dengue virus E glycoprotein are the most efficient blockers of virus adsorption to Vero cells. J Virol 75:7769–7773 [View Article][PubMed]
     [Google Scholar]
  10. Durbin A. P., Whitehead S. S. 2010; Dengue vaccine candidates in development. Curr Top Microbiol Immunol 338:129–143[PubMed]
     [Google Scholar]
  11. Fonseca B. A., Pincus S., Shope R. E., Paoletti E., Mason P. W. 1994; Recombinant vaccinia viruses co-expressing dengue-1 glycoproteins prM and E induce neutralizing antibodies in mice. Vaccine 12:279–285 [View Article][PubMed]
     [Google Scholar]
  12. Goncalvez A. P., Men R., Wernly C., Purcell R. H., Lai C. J. 2004a; Chimpanzee Fab fragments and a derived humanized immunoglobulin G1 antibody that efficiently cross-neutralize dengue type 1 and type 2 viruses. J Virol 78:12910–12918 [View Article][PubMed]
     [Google Scholar]
  13. Goncalvez A. P., Purcell R. H., Lai C. J. 2004b; Epitope determinants of a chimpanzee Fab antibody that efficiently cross-neutralizes dengue type 1 and type 2 viruses map to inside and in close proximity to fusion loop of the dengue type 2 virus envelope glycoprotein. J Virol 78:12919–12928 [View Article][PubMed]
     [Google Scholar]
  14. Gromowski G. D., Barrett A. D. 2007; Characterization of an antigenic site that contains a dominant, type-specific neutralization determinant on the envelope protein domain III (ED3) of dengue 2 virus. Virology 366:349–360 [View Article][PubMed]
     [Google Scholar]
  15. Gromowski G. D., Barrett N. D., Barrett A. D. 2008; Characterization of dengue virus complex-specific neutralizing epitopes on envelope protein domain III of dengue 2 virus. J Virol 82:8828–8837 [View Article][PubMed]
     [Google Scholar]
  16. Gubler D. J. 2002; Epidemic dengue/dengue hemorrhagic fever as a public health, social and economic problem in the 21st century. Trends Microbiol 10:100–103 [View Article][PubMed]
     [Google Scholar]
  17. Guojun W., Wei G., Kedong O., Yi H., Yanfei X., Qingmei C., Yankai Z., Jie W., Hao F. et al. 2008; A novel vaccine targeting gastrin-releasing peptide: efficient inhibition of breast cancer growth in vivo. Endocr Relat Cancer 15:149–159 [View Article][PubMed]
     [Google Scholar]
  18. Halstead S. B. 2003; Neutralization and antibody-dependent enhancement of dengue viruses. Adv Virus Res 60:421–467 [View Article][PubMed]
     [Google Scholar]
  19. Halstead S. B., O’Rourke E. J. 1977; Antibody-enhanced dengue virus infection in primate leukocytes. Nature 265:739–741 [View Article][PubMed]
     [Google Scholar]
  20. Heinz F. X., Stiasny K. 2012; Flaviviruses and flavivirus vaccines. Vaccine 30:4301–4306 [View Article][PubMed]
     [Google Scholar]
  21. Hsu C. T., Ting C. Y., Ting C. J., Chen T. Y., Lin C. P., Whang-Peng J., Hwang J. 2000; Vaccination against gonadotropin-releasing hormone (GnRH) using toxin receptor-binding domain-conjugated GnRH repeats. Cancer Res 60:3701–3705[PubMed]
     [Google Scholar]
  22. Huang K. J., Yang Y. C., Lin Y. S., Huang J. H., Liu H. S., Yeh T. M., Chen S. H., Liu C. C., Lei H. Y. 2006; The dual-specific binding of dengue virus and target cells for the antibody-dependent enhancement of dengue virus infection. J Immunol 176:2825–2832 [View Article][PubMed]
     [Google Scholar]
  23. Hung J. J., Hsieh M. T., Young M. J., Kao C. L., King C. C., Chang W. 2004; An external loop region of domain III of dengue virus type 2 envelope protein is involved in serotype-specific binding to mosquito but not mammalian cells. J Virol 78:378–388 [View Article][PubMed]
     [Google Scholar]
  24. Lee H. C., Butler M., Wu S. C. 2012; Using recombinant DNA technology for the development of live-attenuated dengue vaccines. Enzyme Microb Technol 51:67–72 [View Article][PubMed]
     [Google Scholar]
  25. Leng C. H., Liu S. J., Tsai J. P., Li Y. S., Chen M. Y., Liu H. H., Lien S. P., Yueh A., Hsiao K. N. et al. 2009; A novel dengue vaccine candidate that induces cross-neutralizing antibodies and memory immunity. Microbes Infect 11:288–295 [View Article][PubMed]
     [Google Scholar]
  26. Lin B., Parrish C. R., Murray J. M., Wright P. J. 1994; Localization of a neutralizing epitope on the envelope protein of dengue virus type 2. Virology 202:885–890 [View Article][PubMed]
     [Google Scholar]
  27. Lok S. M., Kostyuchenko V., Nybakken G. E., Holdaway H. A., Battisti A. J., Sukupolvi-Petty S., Sedlak D., Fremont D. H., Chipman P. R. et al. 2008; Binding of a neutralizing antibody to dengue virus alters the arrangement of surface glycoproteins. Nat Struct Mol Biol 15:312–317 [View Article][PubMed]
     [Google Scholar]
  28. Modis Y., Ogata S., Clements D., Harrison S. C. 2003; A ligand-binding pocket in the dengue virus envelope glycoprotein. Proc Natl Acad Sci U S A 100:6986–6991 [View Article][PubMed]
     [Google Scholar]
  29. Modis Y., Ogata S., Clements D., Harrison S. C. 2004; Structure of the dengue virus envelope protein after membrane fusion. Nature 427:313–319 [View Article][PubMed]
     [Google Scholar]
  30. Murphy B. R., Whitehead S. S. 2011; Immune response to dengue virus and prospects for a vaccine. Annu Rev Immunol 29:587–619 [View Article][PubMed]
     [Google Scholar]
  31. Oliphant T., Engle M., Nybakken G. E., Doane C., Johnson S., Huang L., Gorlatov S., Mehlhop E., Marri A. et al. 2005; Development of a humanized monoclonal antibody with therapeutic potential against West Nile virus. Nat Med 11:522–530 [View Article][PubMed]
     [Google Scholar]
  32. Oliphant T., Nybakken G. E., Engle M., Xu Q., Nelson C. A., Sukupolvi-Petty S., Marri A., Lachmi B. E., Olshevsky U. et al. 2006; Antibody recognition and neutralization determinants on domains I and II of West Nile Virus envelope protein. J Virol 80:12149–12159 [View Article][PubMed]
     [Google Scholar]
  33. Rajamanonmani R., Nkenfou C., Clancy P., Yau Y. H., Shochat S. G., Sukupolvi-Petty S., Schul W., Diamond M. S., Vasudevan S. G., Lescar J. 2009; On a mouse monoclonal antibody that neutralizes all four dengue virus serotypes. J Gen Virol 90:799–809 [View Article][PubMed]
     [Google Scholar]
  34. Rey F. A., Heinz F. X., Mandl C., Kunz C., Harrison S. C. 1995; The envelope glycoprotein from tick-borne encephalitis virus at 2 Å resolution. Nature 375:291–298 [View Article][PubMed]
     [Google Scholar]
  35. Roehrig J. T., Bolin R. A., Kelly R. G. 1998; Monoclonal antibody mapping of the envelope glycoprotein of the dengue 2 virus, Jamaica. Virology 246:317–328 [View Article][PubMed]
     [Google Scholar]
  36. Schmitz J., Roehrig J., Barrett A., Hombach J. 2011; Next generation dengue vaccines: a review of candidates in preclinical development. Vaccine 29:7276–7284 [View Article][PubMed]
     [Google Scholar]
  37. Shyu Y. C., Lee T. L., Wen S. C., Chen H., Hsiao W. Y., Chen X., Hwang J., Shen C. K. 2007; Subcellular transport of EKLF and switch-on of murine adult βmaj globin gene transcription. Mol Cell Biol 27:2309–2323 [View Article][PubMed]
     [Google Scholar]
  38. Stiasny K., Kiermayr S., Holzmann H., Heinz F. X. 2006; Cryptic properties of a cluster of dominant flavivirus cross-reactive antigenic sites. J Virol 80:9557–9568 [View Article][PubMed]
     [Google Scholar]
  39. Sukupolvi-Petty S., Austin S. K., Purtha W. E., Oliphant T., Nybakken G. E., Schlesinger J. J., Roehrig J. T., Gromowski G. D., Barrett A. D. et al. 2007; Type- and subcomplex-specific neutralizing antibodies against domain III of dengue virus type 2 envelope protein recognize adjacent epitopes. J Virol 81:12816–12826 [View Article][PubMed]
     [Google Scholar]
  40. Sukupolvi-Petty S., Austin S. K., Engle M., Brien J. D., Dowd K. A., Williams K. L., Johnson S., Rico-Hesse R., Harris E. et al. 2010; Structure and function analysis of therapeutic monoclonal antibodies against dengue virus type 2. J Virol 84:9227–9239 [View Article][PubMed]
     [Google Scholar]
  41. Thullier P., Demangel C., Bedouelle H., Mégret F., Jouan A., Deubel V., Mazié J. C., Lafaye P. 2001; Mapping of a dengue virus neutralizing epitope critical for the infectivity of all serotypes: insight into the neutralization mechanism. J Gen Virol 82:1885–1892[PubMed]
     [Google Scholar]
  42. Volk D. E., Beasley D. W., Kallick D. A., Holbrook M. R., Barrett A. D., Gorenstein D. G. 2004; Solution structure and antibody binding studies of the envelope protein domain III from the New York strain of West Nile virus. J Biol Chem 279:38755–38761 [View Article][PubMed]
     [Google Scholar]
  43. Wan S. W., Lin C. F., Wang S., Chen Y. H., Yeh T. M., Liu H. S., Anderson R., Lin Y. S. 2013; Current progress in dengue vaccines. J Biomed Sci 20:37 [View Article][PubMed]
     [Google Scholar]
  44. Wilder-Smith A., Gubler D. J. 2008; Geographic expansion of dengue: the impact of international travel. Med Clin North Am 92:1377–1390 [View Article][PubMed]
     [Google Scholar]
  45. Yi H., Rong Y., Yankai Z., Wentao L., Hongxia Z., Jie W., Rongyue C., Taiming L., Jingjing L. 2006; Improved efficacy of DNA vaccination against breast cancer by boosting with the repeat beta-hCG C-terminal peptide carried by mycobacterial heat-shock protein HSP65. Vaccine 24:2575–2584 [View Article][PubMed]
     [Google Scholar]
  46. Yu S., Wuu A., Basu R., Holbrook M. R., Barrett A. D., Lee J. C. 2004; Solution structure and structural dynamics of envelope protein domain III of mosquito- and tick-borne flaviviruses. Biochemistry 43:9168–9176 [View Article][PubMed]
     [Google Scholar]
  47. Zhang Y., Corver J., Chipman P. R., Zhang W., Pletnev S. V., Sedlak D., Baker T. S., Strauss J. H., Kuhn R. J., Rossmann M. G. 2003; Structures of immature flavivirus particles. EMBO J 22:2604–2613 [View Article][PubMed]
     [Google Scholar]
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Production of a neutralizing antibody against envelope protein of dengue virus type 2 using the linear array epitope technique
J. Gen. Virol. 95, 2155 (2014); https://doi.org/10.1099/vir.0.062562-0
/content/journal/jgv/10.1099/vir.0.062562-0
/content/journal/jgv/10.1099/vir.0.062562-0
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