Isolation of recombinant antibodies directed against surface proteins of Clostridium difficile

Shirvan, A. N. and Aitken, R. (2016) Isolation of recombinant antibodies directed against surface proteins of Clostridium difficile. Brazilian Journal of Microbiology, 47(2), pp. 394-402. 26991284. (doi: 10.1016/j.bjm.2016.01.017) (PMID:PMC4874623)

Abstract

Clostridium difficile has emerged as an increasingly important nosocomial pathogen and the prime causative agent of antibiotic-associated diarrhoea and pseudomembranous colitis in humans. In addition to toxins A and B, immunological studies using antisera from patients infected with C. difficile have shown that a number of other bacterial factors contribute to the pathogenesis, including surface proteins, which are responsible for adhesion, motility and other interactions with the human host. In this study, various clostridial targets, including FliC, FliD and cell wall protein 66, were expressed and purified. Phage antibody display yielded a large panel of specific recombinant antibodies, which were expressed, purified and characterised. Reactions of the recombinant antibodies with their targets were detected by enzyme-linked immunosorbent assay; and Western blotting suggested that linear rather than conformational epitopes were recognised. Binding of the recombinant antibodies to surface-layer proteins and their components showed strain specificity, with good recognition of proteins from C. difficile 630. However, no reaction was observed for strain R20291—a representative of the 027 ribotype. Binding of the recombinant antibodies to C. difficile M120 extracts indicated that a component of a surface-layer protein of this strain might possess immunoglobulin-binding activities. The recombinant antibodies against FliC and FliD proteins were able to inhibit bacterial motility.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Aitken, Professor Robert
• Authors: Shirvan, A. N., and Aitken, R.
• College/School: College of Medical Veterinary and Life Sciences
• Journal Name: Brazilian Journal of Microbiology
• Publisher: Sociedade Brasileira de Microbiologia
• ISSN: 1517-8382
• ISSN (Online): 1678-4405
• Published Online: 02 March 2016
• Copyright Holders: Copyright © 2016 Elsevier
• First Published: First published in Brazilian Journal of Microbiology 47(2):394-402
• Publisher Policy: Reproduced under a Creative Commons License