This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Leung, P. H. M.
Right arrow Articles by Yam, W. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Leung, P. H. M.
Right arrow Articles by Yam, W. C.

 Previous Article  |  Next Article 

Clinical and Diagnostic Laboratory Immunology, May 2002, p. 687-692, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.687-692.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Polyclonal Antibodies to Glutathione S-Transferase- Verotoxin Subunit A Fusion Proteins Neutralize Verotoxins

P. H. M. Leung,1 J. S. M. Peiris,1 W. W. S. Ng,2 and W. C. Yam1*

Department of Microbiology, Queen Mary Hospital,1 School of Professional and Continual Education, The University of Hong Kong, Hong Kong, Special Administrative Region, People's Republic of China2

Received 19 November 2001/ Returned for modification 8 January 2002/ Accepted 20 February 2002

The A1 subunits of verotoxin-1 (VT1) and VT2 genes were cloned into pGEX-4T-2 for the expression of glutathione S-transferase (GST) fusion proteins. The N-terminal and the transmembrane regions of the A1 subunits were excluded from the constructs in order to increase the product yields. Polyclonal anti-VT1A1 and anti-VT2A1 antibodies were produced by immunizing rabbits with GST-VT1A1 and GST-VT2A1 fusion proteins, respectively. The antibodies were tested for their ability to neutralize active toxins from 45 VT-producing Escherichia coli (VTEC) strains. The antibodies had significantly high neutralizing activities against their homologous toxins. The average percentages of neutralization of VT1 by anti-GST-VT1A1 and anti-GST-VT2A1 were 76.7% ± 7.9% and 3.6% ± 2.3%, respectively, and those of VT2 were 1.7% ± 2.3% and 82.5% ± 13.9%, respectively. VT2 variant toxin was neutralized by anti-GST-VT2A1, with cross neutralization being a possible consequence of sequence homology between VT2 and a VT2 variant. To our knowledge, this is the first report on the production of polyclonal antibodies from GST-VT fusion proteins. The antibodies were shown to exhibit specific toxin neutralizing activities and may be useful for immunological diagnosis of VTEC infections.

* Corresponding author. Mailing address: Department of Microbiology, Queen Mary Hospital, The University of Hong Kong, Hong Kong SAR, China. Phone: 852 28554821. Fax: 852 28551241. E-mail: wcyam{at}hkucc.hku.hk.

Clinical and Diagnostic Laboratory Immunology, May 2002, p. 687-692, Vol. 9, No. 3
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.3.687-692.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Leung, P. H. M., Peiris, J. S. M., Ng, W. W. S., Robins-Browne, R. M., Bettelheim, K. A., Yam, W. C. (2003). A Newly Discovered Verotoxin Variant, VT2g, Produced by Bovine Verocytotoxigenic Escherichia coli. Appl. Environ. Microbiol. 69: 7549-7553 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»