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Open Access 01-12-2021 | Escherichia Coli | Research article

Isolation and characterizations of a novel recombinant scFv antibody against exotoxin A of Pseudomonas aeruginosa

Authors: Zahra Shadman, Safar Farajnia, Mohammad Pazhang, Mohammadreza Tohidkia, Leila Rahbarnia, Saeed Najavand, Sayna Toraby

Published in: BMC Infectious Diseases | Issue 1/2021

Abstract

Background

Pseudomonas aeruginosa is the leading cause of nosocomial infections, especially in people with a compromised immune system. Targeting virulence factors by neutralizing antibodies is a novel paradigm for the treatment of antibiotic-resistant pseudomonas infections. In this respect, exotoxin A is one of the most potent virulence factors in P. aeruginosa. The present study was carried out to identify a novel human scFv antibody against the P. aeruginosa exotoxin A domain I (ExoA-DI) from a human scFv phage library.

Methods

The recombinant ExoA-DI of P. aeruginosa was expressed in E. coli, purified by Ni-NTA column, and used for screening of human antibody phage library. A novel screening procedure was conducted to prevent the elimination of rare specific clones. The phage clone with high reactivity was evaluated by ELISA and western blot.

Results

Based on the results of polyclonal phage ELISA, the fifth round of biopanning leads to the isolation of several ExoA-DI reactive clones. One positive clone with high affinity was selected by monoclonal phage ELISA and used for antibody expression. The purified scFv showed high reactivity with the recombinant domain I and full-length native exotoxin A.

Conclusions

The purified anti-exotoxin A scFv displayed high specificity against exotoxin A. The human scFv identified in this study could be the groundwork for developing a novel therapeutic agent to control P. aeruginosa infections.

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Title: Isolation and characterizations of a novel recombinant scFv antibody against exotoxin A of Pseudomonas aeruginosa
Authors: Zahra Shadman
Safar Farajnia
Mohammad Pazhang
Mohammadreza Tohidkia
Leila Rahbarnia
Saeed Najavand
Sayna Toraby
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Escherichia Coli
Pseudomonas
Pseudomonas Aeruginosa
Published in: BMC Infectious Diseases / Issue 1/2021
Electronic ISSN: 1471-2334
DOI: https://doi.org/10.1186/s12879-021-05969-0

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Isolation and characterizations of a novel recombinant scFv antibody against exotoxin A of Pseudomonas aeruginosa

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Conclusions

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