European Journal of Microbiology and Immunology
Volume/Issue:
Volume 4: Issue 1
Recombinant human antibody fragment against tetanus toxoid produced by phage display
Authors:
B. Neelakantam Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India
Department of Biotechnology, Acharya Nagarjuna University, Guntur, 522510, India

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N. V. Sridevi Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India

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A. M. Shukra Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India

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P. Sugumar Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India

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S. Samuel Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India

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Dr. Lingala Rajendra PhD Research and Development Center, Indian Immunologicals Limited, Hyderabad, Gachibowli, 500032, India

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Pages:
45–55
Online Publication Date:
01 Mar 2014
Publication Date:
01 Mar 2014
Article Category:
Research Article
DOI:
https://doi.org/10.1556/eujmi.4.2014.1.4
Keywords:
Fabs; TT; flocculation; ELISA; phage
Restricted access

Abstract

Phage display technology is a powerful in vitro method for the identification of specific monoclonal antibodies (antibody fragments) to an antigenic target and allows the rapid generation and selection of high affinity, fully human antibodies directed toward any disease target appropriate for antibody therapy. In the present study, we exploited the phage display technology for the selection of an antigen binding fragment (Fabs) toward tetanus toxoid using human naïve phage antibody library constructed from peripheral blood lymphocytes of naïve human donors. The phages displaying Fab were subjected to three rounds of bio-panning with tetanus toxoid as antigen on a solid phase. The high affinity antibody fragments were expressed in HB2151 strain of Escherichia coli and purified by immobilized metal affinity chromatography. The binding activity and specificity of the antibody fragment was established by its reactivity toward tetanus toxoid and non-reactivity toward other related toxins as determined by enzyme-linked immunosorbent assay and immunoblot analysis. The selected Fab fragment forming the antigen-binding complexes with the toxoid in flocculation assay indicates that the Fab may have a potential neutralizing ability toward antigen.

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Cover European Journal of Microbiology and Immunology
European Journal of Microbiology and Immunology
Print ISSN:
2062-509X
Online ISSN:
2062-8633
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Editor(s)-in-Chief: Dunay, Ildiko Rita, Prof. Dr. Pharm, Dr. rer. nat., University of Magdeburg, Germany

Editor(s)-in-Chief: Heimesaat, Markus M., Prof. Dr. med., Charité - University Medicine Berlin, Germany

Editorial Board

  • Berit Bangoura, Dr. DVM. PhD,  University of Wyoming, USA
  • Stefan Bereswill, Prof. Dr. rer. nat., Charité - University Medicine Berlin, Germany
  • Dunja Bruder, Prof. Dr. rer. nat., University of Magdeburg, Germany
  • Jan Buer, Prof. Dr. med., University of Duisburg, Germany
  • Edit Buzas, Prof. Dr. med., Semmelweis University, Hungary
  • Renato Damatta, Prof. PhD, UENF, Brazil
  • Maria Deli, MD, PhD, DSc, Biological Research Center, HAS, Hungary
  • Olgica Djurković-Djaković, Prof. Phd, University of Belgrade, Serbia
  • Jean-Dennis Docquier, Prof. Dr. med., University of Siena, Italy
  • Zsuzsanna Fabry, Prof. Phd, University of Washington, USA
  • Ralf Ignatius, Prof. Dr. med., Charité - University Medicine Berlin, Germany
  • Achim Kaasch, Prof. Dr. med., Otto von Guericke University Magdeburg, Germany
  • Oliver Liesenfeld, Prof. Dr. med., Inflammatix, USA
  • Matyas Sandor, Prof. PhD, University of Wisconsin, USA
  • Ulrich Steinhoff, Prof. PhD, University of Marburg, Germany
  • Michal Toborek, Prof. PhD, University of Miami, USA
  • Susanne A. Wolf, PhD, MDC-Berlin, Germany

 

Dr. Dunay, Ildiko Rita
Magdeburg, Germany
E-mail: ildiko.dunay@med.ovgu.de

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European Journal of Microbiology and Immunology
Language English
Size A4
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Foundation		 2011
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per Year		 1
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per Year		 4
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ISSN 2062-509X (Print)
ISSN 2062-8633 (Online)

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