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Ribosome display of combinatorial antibody libraries derived from mice immunised with heat-killed Xylella fastidiosa and the selection of MopB-specific single-chain antibodies

Azizi, Armaghan and Arora, Arinder and Markiv, Anatoliy and Lampe, David J. and Miller, Thomas and Kang, Angray S. (2012) Ribosome display of combinatorial antibody libraries derived from mice immunised with heat-killed Xylella fastidiosa and the selection of MopB-specific single-chain antibodies. Applied and Environmental Microbiology, 78 (8). pp. 2638-2647. ISSN 0099-2240

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Official URL: http://dx.doi.org/10.1128/AEM.07807-11

Abstract

Pierce's disease is a devastating lethal disease of Vitus vinifera grapevines caused by the bacterium Xylella fastidiosa. There is no cure for Pierce's disease and control is achieved predominantly by suppressing transmission of the glassy winged sharpshooter insect vector. We present a simple robust approach for the generation of panels of recombinant single chain antibodies against the surface exposed elements of X. fastidiosa that may have potential use in diagnosis and/or disease transmission blocking studies. In vitro combinatorial antibody ribosome display libraries were assembled from immunoglobulin transcripts rescued from the spleens of mice immunized with heat-killed X. fastidiosa. The libraries were used in a single round of selection against an outer-membrane protein MopB, resulting in the isolation of a panel of recombinant antibodies. The potential use of selected anti-MopB antibodies was demonstrated by the successful application of the 4XfMopB3 antibody in an ELISA, western blot and immunofluorescence assay. These immortalised in vitro recombinant single chain antibody libraries generated against heat killed X. fastidiosa are a resource for the Pierce's disease research community that may be readily accessed for the isolation of antibodies against a plethora of X. fastidiosa surface exposed antigenic molecules.

Item Type:Article
Research Community:University of Westminster > Life Sciences, School of
ID Code:10315
Deposited On:21 Feb 2012 11:48
Last Modified:18 Oct 2012 11:37

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