Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Production of a phage-displayed single chain variable fragment antibody against infectious bursal disease virus

Amir Shabdini Pashaki¹, Mohammad Reza Safarnejad² , Amir Hossein Asgari Safdar³, Hosein Safarpour⁴, Meisam Tabatabaie⁵

¹Department of Animal Science, Science and Research Branch, Islamic Azad University, Tehran, Iran; ²Iranian Research Institute of Plant Protections, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran; ³Young Researchers and Elite Club, Baft Branch, Islamic Azad University, Baft, Iran; ⁴Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran; ⁵Microbial biotechnology Department, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

For correspondence:- Mohammad Safarnejad Email: mrsafarnejad@yahoo.com Tel:+982122403012

Accepted: 27 November 2017 Published: 29 December 2017

Citation: Pashaki AS, Safarnejad MR, Safdar AH, Safarpour H, Tabatabaie M. Production of a phage-displayed single chain variable fragment antibody against infectious bursal disease virus. Trop J Pharm Res 2017; 16(12):2801-2809 doi: 10.4314/tjpr.v16i12.3

© 2017 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To develop specific single chain variable fragments (scFv) against infectious bursal disease virus (IBDV) via phage display technology.

Methods: Purified viruses were initially applied for iterative panning rounds of scFv phage display libraries. The binding ability of the selected scFv antibody fragments against the IBDV particles was analyzed by indirect enzyme-linked immunosorbent assay (ELISA) followed by blotting assays. Three-dimensional (3D) structure of the selected scFv antibody fragment and VP3 protein were predicted through in silico analysis. Structural characterization of the antibody-antigen complexes was carried out by computational docking analysis.

Results: The serological results obtained from the ELISA and blotting analysis showed that the selected clones produced specific scFv antibody fragments that were capable of effectively detecting infectious bursal disease (IBD) in the infected animal tissue. Biodiversity analysis by BstNI finger printing and nucleotide sequencing revealed that there was no major difference in nucleotide sequences of the selected clones. Further analysis demonstrated that this recombinant fragment of the antibody was able to bind to VP3 structural protein of IBDV with a molecular weight of ~30 kDa. Molecular docking results revealed that the binding energy of scFv to IBDV-VP3 was 545 kj/mol.

Conclusion: The developed scFv antibody fragments possess great potentials for the diagnosis of IBD. The findings of the present study confirm the feasibility of using phage display technology for rapid production of antibodies against IBD diseases by applying naïve scFv libraries

Keywords: Antibody, Molecular docking, Phage display technology, Single chain variable fragments