Immunologic Research

, Volume 62, Issue 3, pp 325–340 | Cite as

A repertoire of high-affinity monoclonal antibodies specific to S. typhi: as potential candidate for improved typhoid diagnostic

  • Chandresh Sharma
  • Anurag Sankhyan
  • Tarang Sharma
  • Naeem Khan
  • Susmita Chaudhuri
  • Niraj Kumar
  • Shinjini Bhatnagar
  • Navin Khanna
  • Ashutosh Tiwari


Typhoid fever is a significant global health problem with highest burden on the developing world. The severity of typhoid is often underestimated, and currently available serological diagnostic assays are inadequate due to lack in requisite sensitivity and specificity. This underlines an absolute need to develop a reliable and accurate diagnostics that would benefit long-term disease control and treatment and to understand the real disease burden. Here, we have utilized flagellin protein of S. typhi that is surface accessible, abundantly expressed, and highly immunogenic, for developing immunodiagnostic tests. Flagellin monomers are composed of conserved amino-terminal and carboxy-terminal, and serovar-specific middle region. We have generated a panel of murine monoclonal antibodies (mAbs) against the middle region of flagellin, purified from large culture of S. typhi to ensure its native conformation. These mAbs showed unique specificity and very high affinity toward S. typhi flagellin without showing any cross-reactivity with other serovars. Genetic analysis of mAbs also revealed high frequency of somatic mutation due to antigenic selection process across variable region to achieve high binding affinity. These antibodies also displayed stable binding in stringent reaction conditions for antigen–antibody interactions, like DMSO, urea, KSCN, guanidinium HCl, and extremes of pH. One of the mAbs potentially reversed the TLR5-mediated immune response, in vitro by inhibiting TLR5–flagellin interaction. In our study, binding of these mAbs to flagellin, with high affinity, present on bacterial surface, as well as in soluble form, validates their potential use in developing improved diagnostics with significantly higher sensitivity and specificity.


Flagellin S.typhi Typhoid fever Monoclonal antibodies TLR5 Typhoid diagnostics 



The authors gratefully acknowledge Dr. Ayub Qadri (National Institute of Immunology, New Delhi, India) and Dr. Bhabatosh Das (Translational Health Science and Technology Institute), for providing Salmonella strains. This study was supported by grant from Department of Biotechnology, Government of India, to Centre for Bio-design and Diagnostics, Translational Health Science and Technology Institute, and the core grant of Translational Health Science and Technology Institute to AT. C S was supported by Innovation Award from Centre for Bio-design and Diagnostics.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

All procedures performed in this study involving animals were in accordance with the ethical standards of the institution or practice at the International Centre for Genetic Engineering and Biotechnology, New Delhi, India.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Chandresh Sharma
    • 1
  • Anurag Sankhyan
    • 1
  • Tarang Sharma
    • 1
  • Naeem Khan
    • 2
  • Susmita Chaudhuri
    • 1
  • Niraj Kumar
    • 1
  • Shinjini Bhatnagar
    • 3
  • Navin Khanna
    • 4
  • Ashutosh Tiwari
    • 1
  1. 1.Centre for Bio-design and DiagnosticsTranslational Health Science and Technology InstituteFaridabadIndia
  2. 2.National Institute of ImmunologyNew DelhiIndia
  3. 3.Pediatric Biology CenterTranslational Health Science and Technology InstituteFaridabadIndia
  4. 4.International Centre for Genetic Engineering and BiotechnologyNew DelhiIndia

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