Type

Journal Article

Authors

Richard O'Kennedy
John Quinn
Paul Leonard
Stephen Hearty

Subjects

Microbiology

Topics
isolation purification antigens bacterial bacterial proteins recombinant proteins immunology female prevention control chemistry enzyme linked immunosorbent assay listeria monocytogenes surface plasmon resonance genetics mice inbred balb c antibodies monoclonal epitopes sensitivity and specificity biosynthesis biosensing techniques listeriosis animals internalin protein bacteria blotting western food microbiology mice microscopy atomic force

Production, characterisation and potential application of a novel monoclonal antibody for rapid identification of virulent Listeria monocytogenes. (2005)

Abstract A panel of hybridomas was produced using intact Listeria monocytogenes serotype 1/2a cells as the immunogen. An IgG2a monoclonal antibody (mAb) 'mAb2B3' was isolated that reacted with L. monocytogenes but not with a representative panel of related Listeria spp. and non-Listeria spp. Binding activity was greatest against L. monocytogenes serotype 1/2a and was significantly enhanced when cells were prepared in Listeria enrichment broth (LEB). The reactive epitope was deduced, by immunoblot analysis, to be a surface localised protein of approximately 80 kilodaltons (kDa), putatively assumed to be internalin A (InlA). Recombinant InlA protein was subsequently expressed in Escherischia coli. When crude E. coli cell lysates were subjected to immunoblot analysis, it was demonstrated that the mAb bound specifically to the heterologously expressed recombinant InlA protein, thus confirming the specificity of the mAb. The mAb was further evaluated in a series of enzyme-linked-immunosorbent assay (ELISA)-based formats and in a surface plasmon resonance (SPR)-based biosensor platform. Both configurations were capable of differential identification of virulent L. monocytogenes at concentrations greater than or equal to 1x10(7) cells/ml. Notwithstanding the apparent insensitivity, the results indicate that InlA could be exploited as a marker for highly specific confirmatory identification of pathogenic L. monocytogenes following primary enrichment of suspect food samples, using the anti-InlA antibody 'mAb2B3', described herein.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Richard O'Kennedy, John Quinn, Paul Leonard, Stephen Hearty

Experts in our system

1
R O'Kennedy
Dublin City University
Total Publications: 197
 
2
Paul Leonard
Dublin City University
Total Publications: 27
 
3
Stephen Hearty
Dublin City University
Total Publications: 27