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

BMC Infectious Diseases

Sayna Toraby¹  Saeed Najavand¹  Mohammad Pazhang²  Safar Farajnia³  Zahra Shadman⁴  Leila Rahbarnia⁵  Mohammadreza Tohidkia⁶ 

[1] Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;Department of Cellular and Molecular Biology, Faculty of Science Faculty, Azarbaijan Shahid Madani University, Tabriz, Iran;Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;Department of Cellular and Molecular Biology, Faculty of Science Faculty, Azarbaijan Shahid Madani University, Tabriz, Iran;Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;Nanotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran;

关键词: Phage display;    Pseudomonas aeruginosa;    Exotoxin A;    Human single chain antibody;   

DOI  :  10.1186/s12879-021-05969-0

来源: Springer

【 摘 要 】

BackgroundPseudomonas 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.MethodsThe 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.ResultsBased 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.ConclusionsThe 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.

【 授权许可】

CC BY