Archives of Virology

, Volume 160, Issue 11, pp 2749–2761 | Cite as

Development of novel antibodies against non-structural proteins nsP1, nsP3 and nsP4 of chikungunya virus: potential use in basic research

  • Sameer Kumar
  • Prabhudutta Mamidi
  • Abhishek Kumar
  • Itishree Basantray
  • Umarani Bramha
  • Anshuman Dixit
  • Prasanta Kumar Maiti
  • Sujay Singh
  • Amol Ratnakar Suryawanshi
  • Subhasis Chattopadhyay
  • Soma ChattopadhyayEmail author
Original Article


Chikungunya virus (CHIKV) has reemerged recently as an important pathogen, causing several large epidemics worldwide. This necessitates the development of better reagents to understand its biology and to establish effective and safe control measures. The present study describes the development and characterization of polyclonal antibodies (pAbs) against synthetic peptides of CHIKV non-structural proteins (nsPs; nsP1, nsP3 and nsP4). The reactivity of these pAbs was demonstrated by ELISA and Western blot. Additionally, in vitro infection studies in a mammalian system confirmed that these pAbs are highly sensitive and specific for CHIKV nsPs, as these proteins were detected very early during viral replication. Homology analysis of the selected epitope sequences revealed that they are conserved among all of the CHIKV strains of different genotypes, while comparison with other alphavirus sequences showed that none of them are 100 % identical to the epitope sequences (except Onyong-nyong and Igbo Ora viruses, which show 100 % identity to the nsP4 epitope). Interestingly, two different forms of CHIKV nsP1 and three different forms of nsP3 were detected in Western blot analysis during infection; however, further experimental investigations are required to confirm their identity. Also, the use of these antibodies demonstrated faster and enhanced expression profiles of all CHIKV nsPs in 2006 Indian outbreak strains when compared to the CHIKV prototype strain, suggesting the epidemic potential of the 2006 isolate. Accordingly, it can be suggested that the pAbs reported in this study can be used as sensitive and specific tools for experimental investigations of CHIKV replication and infection.


Vero Cell Semliki Forest Virus Chikungunya Virus CHIKV Infection nsP4 Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank Dr. Manmohan Parida for kindly providing the virus strains (S-27 and DRDE-06), CHIKV polyclonal antibody, and Vero cell lines. This work was supported by Department of Biotechnology, Ministry of Science and Technology, Govt. of India vide grant no. BT/PR13118/GBD/27/186/2009, BT/PR15173/GBD/27/356/2011 and by Council of Scientific and Industrial Research (CSIR), Ministry of Science and Technology, Govt. of India vide Project No. 37(1542)/12/EMR-II).


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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Sameer Kumar
    • 1
  • Prabhudutta Mamidi
    • 1
  • Abhishek Kumar
    • 1
  • Itishree Basantray
    • 1
  • Umarani Bramha
    • 1
  • Anshuman Dixit
    • 1
  • Prasanta Kumar Maiti
    • 2
  • Sujay Singh
    • 2
  • Amol Ratnakar Suryawanshi
    • 1
  • Subhasis Chattopadhyay
    • 3
  • Soma Chattopadhyay
    • 1
    Email author
  1. 1.Infectious Disease BiologyInstitute of Life SciencesBhubaneswarIndia
  2. 2.Imgenex India Pvt. Ltd.BhubaneswarIndia
  3. 3.School of Biological SciencesNational Institute of Science Education and ResearchBhubaneswarIndia

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