ISSN: 0973-7510

E-ISSN: 2581-690X

Chidanand A. Rabinal1 , Narayan Moger1, Satish Verma1, P. U. Krishnaraj2 and K. N. Chandrashekara3
1Department of Biotechnology, College of Agricultural Dharwad, University of Agricultural Sciences, Dharwad, Karnataka, India.
2Department of Agricultural Microbiology, College of Agricultural Vijayapur, University of Agriculutral Sciences, Dharwad, Karnataka, India.
3Department of Plant Physiology and Biotechnology UPASI TRI TRF, Valparai, Tamil Nadu, India.
J Pure Appl Microbiol. 2015;9(2):1039-1046
© The Author(s). 2015
Received: 13/04/2015 | Accepted: 09/05/2015 | Published: 30/06/2015
Abstract

Phage display technology was used to produce recombinant monoclonal antibody against Bacillus thuringiensis crystalline protein (Cry2B) from Tomlinson I antibody library. The polyclonal ELISA had revealed the fourth round of biopan shown highest specificity (1.480) and 92.5 folds greater than that of control. Total forty-five monoclonal antibodies were screened from fourth round of biopanning against Cry2B. Among forty-five clones, pscFvCry2B19 and pscFvCry2B43 had highest specificity against target. Further, the clones pscFvCry2B19 and pscFvCry2B43 were validated for their cross reactivity with other Cry proteins and the clones were sequenced with LMB3 forward and pHEN reverse primers. The scFv fragments of pscFvCry2B19 and pscFvCry2B43 were of 746 bp and 747 bp long respectively and almost similar to each other at DNA and amino acid level except at C’ terminal end. The produced scFv monoclones have the potential to detect the Cry2B antigen.

Keywords

Phage display technology, scFv,  Biopanning, ELISA, Cry2B antigen, Bacillus thuringiensis

Article Metrics

Article View: 768

Share This Article

© The Author(s) 2015. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.