AIMS Microbiology, 2017, 3(2): 248-266. doi: 10.3934/microbiol.2017.2.248.

Research article

Export file:

Format

  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text

Content

  • Citation Only
  • Citation and Abstract

Evaluation of antibody response against recombinant domain III proteins of dengue virus type 1 and 2

Bioprocess Scale up Facility, Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, India

Dengue, a mosquito borne viral disease caused by dengue virus has emerged as a major health problem during the last few decades. The envelope domain III (DIII) protein of dengue virus is highly immunogenic and capable of inducing neutralizing antibodies against wild-type dengue virus. The envelope domain III protein is a potential subunit vaccine candidate as well as a diagnostic reagent for dengue. This report describes the high yield production and immunogenicity of recombinant DIII proteins of dengue virus type 1 and 2. The subunit DIII proteins were produced in Escherichia coli using batch and fed-batch fermentation process. Immobilized metal affinity chromatography was used to capture DIII proteins of dengue virus type 1 and 2. The purified proteins were refolded by diafiltration to achieve biologically active proteins. After fed-batch fermentation, the recombinant E. coli resulted in purified DIII proteins of about 10.06 mg and 47.70 mg per gram of dry cell weight for recombinant dengue virus type 1 and 2 respectively with more than 95% purity. Biological function of the purified DIII proteins were confirmed by their ability to generate DIII specific antibodies in mice. The DIII antigens in combination with adjuvant resulted antibody endpoint titers of 1:64,000 and 1:1,28,000 for recombinant dengue virus type 1 and 2 respectively. These findings establish that the DIII proteins in combination with adjuvant are immunogenic, which suggests that refolded and purified DIII proteins can be a potential vaccine candidates.
  Figure/Table
  Supplementary
  Article Metrics

Keywords fermentation; scale up; purification; vaccine; ELISA; immunogenicity

Citation: Nagesh K Tripathi, Ambuj Shrivastava. Evaluation of antibody response against recombinant domain III proteins of dengue virus type 1 and 2. AIMS Microbiology, 2017, 3(2): 248-266. doi: 10.3934/microbiol.2017.2.248

References

  • 1. Guzman MG, Harris E (2015) Dengue. Lancet 385: 453–465.    
  • 2. Halstead SB (2007) Dengue. Lancet 370: 1644–1652.    
  • 3. Chiang CY, Huang MH, Hsieh CH, et al. (2012) Dengue-1 envelope protein domain III along with PELC and CpG oligodeoxynucleotides synergistically enhances immune responses. PLoS Negl Trop Dis 6: e1645.    
  • 4. Guzman MG, Hermida L, Bernardo L, et al. (2010) Domain III of the envelope protein as a dengue vaccine target. Expert Rev Vaccines 9: 137–147.    
  • 5. Cardoso SA, Paixao VF, Oliveira MD, et al. (2013) Dengue 1 envelope protein domain III produced in Pichia pastoris: potential use for serological diagnosis. Protein Expr Purif 92: 9–13.    
  • 6. Izquierdo A, Garcia A, Lazo L, et al. (2014) A tetravalent dengue vaccine containing a mix of domain III-P64k and domain III-capsid proteins induces a protective response in mice. Arch Virol 159: 2597–2604.    
  • 7. Suzarte E, Marcos E, Gil L, et al. (2014) Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus. Arch Virol 159: 1629–1640.    
  • 8. Tan LCM, Chua AJS, Goh LSL, et al. (2010) Rapid purification of recombinant dengue and West Nile virus envelope Domain III proteins by metal affinity membrane chromatography. Protein Expr Purif 74: 129–139.    
  • 9. Babu JP, Pattnaik P, Gupta N, et al. (2008) Immunogenicity of a recombinant envelope domain III protein of dengue virus type-4 with various adjuvants in mice. Vaccine 26: 4655–4663.    
  • 10. Niu G, Pang Z, Guan C, et al. (2015) Dengue virus envelope domain III protein based on a tetravalent antigen secreted from insect cells: Potential use for serological diagnosis. Virus Res 201: 73–78.    
  • 11. Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27: 297–306.    
  • 12. Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5: 172.
  • 13. Sahdev S, Khattar SK, Saini KS (2008) Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies. Mol Cell Biochem 307: 249–264.
  • 14. Huang, CJ, Lin H, Yang X (2012) Industrial production of recombinant therapeutics in E. coli and its recent advancements. J Ind Microbiol Biotechnol 39: 383–399.
  • 15. Khalilzadeh R, Mohammadian MJ, Bahrami A, et al. (2008) Process development for production of human granulocyte-colony stimulating factor by high cell density cultivation of recombinant Escherichia coli. J Ind Microbiol Biotechnol 35: 1643–1650.    
  • 16. Fong BA, Wood DW (2010) Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentation. Microb Cell Fact 9: 77.    
  • 17. Rathore AS, Bade P, Joshi V, et al. (2013) Refolding of biotech therapeutic proteins expressed in bacteria: review. J Chem Technol Biotechnol 88: 1794–1806.    
  • 18. Singh SM, Panda AK (2005) Solubilization and refolding of bacterial inclusion body proteins. J Biosci Bioeng 99: 303–310.    
  • 19. Tripathi NK (2016) Production and purification of recombinant proteins from Escherichia coli. ChemBioEng Rev 3: 116–133.    
  • 20. Wang Y, Ren W, Gao D, et al. (2015) One-step refolding and purification of recombinant human tumor necrosis factor-α (rhTNF-α) using ion-exchange chromatography. Biomed Chromatogr 29: 305–311.    
  • 21. Sereikaite J, Statkute A, Morkunas M, et al. (2007) Production of recombinant mink growth hormone in E. coli. Appl Microbiol Biotechnol 74: 316–323.    
  • 22. Wang C, Wang L, Geng, X (2009) Optimization of refolding with simultaneous purification of recombinant human granulocyte colony-stimulating factor from Escherichia coli by immobilized metal ion affinity chromatography. Biochem Eng J 43: 197–202.    
  • 23. Tripathi NK, Biswal KC, Rao PV (2015) Scaling up of recombinant dengue virus type 3 envelope domain III protein production from Escherichia coli. Ind Biotechnol 11: 331–337.    
  • 24. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.    
  • 25. Verma SK, Kumar S, Gupta N, et al. (2009) Bacterially expressed recombinant envelope protein domain III of Japanese encephalitis virus (rJEV-DIII) elicits Th1 type of immune response in BALB/c mice. Vaccine 27: 6905–6909.    
  • 26. Tripathi NK, Saxena P, Sathyaseelan, et al. (2008). Production of recombinant dengue virus type 2 envelope domain III protein in Escherichia coli. Ind Chem Engr 50: 194–204.
  • 27. Nausch H, Huckauf J, Koslowski R, et al. (2013) Recombinant production of human interleukin 6 in Escherichia coli. PLoS One 8: e54933.    
  • 28. Manderson D, Dempster R, Chisti Y (2006) Recombinant vaccine against hydatidosis: production of the antigen in E. coli. J Ind Microbiol Biotechnol 33: 173–182.    
  • 29. Volonte F, Piubelli L, Pollegioni L (2011) Optimizing HIV-1 protease production in Escherichia coli as fusion protein. Microb Cell Fact 10: 53.    
  • 30. Bell BA, Wood JF, Bansal R, et al. (2009) Process development for the production of an E. coli produced clinical grade recombinant malaria vaccine for Plasmodium vivax. Vaccine 27: 1448–1453.
  • 31. Buckland BC (2005) The process development challenge for a new vaccine. Nat Med 11: S16–S19.    
  • 32. Babaeipour V, Shojaosadati SA, Khalilzadeh R, et al. (2010) Enhancement of human gamma-interferon production in recombinant E. coli using batch cultivation. Appl Biochem Biotechnol 160: 2366–2376.
  • 33. Lim HK, Jung KH, Park DH, et al. (2000) Production characteristics of interferon-α using an l-arabinose promoter system in a high-cell-density culture. Appl Microbiol Biotechnol 53: 201–208.    
  • 34. Batra G, Gurramkonda C, Nemani SK, et al. (2010) Optimization of conditions for secretion of dengue virus type 2 envelope domain III using Pichia pastoris. J Biosci Bioeng 110: 408–414.35. Valdes I, Bernardo L, Gil L, et al. A novel fusion protein domain III-capsid from dengue-2, in a highly aggregated form, induces a functional immune response and protection in mice. Virology 394: 249–258.    
  • 35. Valdes I, Bernardo L, Gil L, et al. A novel fusion protein domain III-capsid from dengue-2, in a highly aggregated form, induces a functional immune response and protection in mice. Virology 394: 249–258.

 

This article has been cited by

  • 1. Nagesh K. Tripathi, Ambuj Shrivastava, , Nanoscale Fabrication, Optimization, Scale-Up and Biological Aspects of Pharmaceutical Nanotechnology, 2018, 133, 10.1016/B978-0-12-813629-4.00004-8
  • 2. Nagesh K. Tripathi, Divyanshi Karothia, Ambuj Shrivastava, Swati Banger, Jyoti S. Kumar, Enhanced production and immunological characterization of recombinant West Nile virus envelope domain III protein, New Biotechnology, 2018, 46, 7, 10.1016/j.nbt.2018.05.002
  • 3. Nagesh K. Tripathi, Ambuj Shrivastava, Recent Developments in Recombinant Protein–Based Dengue Vaccines, Frontiers in Immunology, 2018, 9, 10.3389/fimmu.2018.01919

Reader Comments

your name: *   your email: *  

Copyright Info: 2017, Nagesh K Tripathi, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Copyright © AIMS Press All Rights Reserved