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A fully recombinant ELISA using in vivo biotinylated antibody fragments for the detection of potato leafroll virus

Lookup NU author(s): Dr Ahmad Al-Mrabeh


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A recombinant antibody fusion protein, V3HCL, which was shown previously to have specific reactivity for potato leafroll virus (PLRV), was labeled with biotin using standard chemical coupling procedures and by an in vivo method. The in vivo method proved superior giving reproducible V3HCL-biotin preparations. A fully recombinant ELISA was devised incorporating V3HCL, V3HCL-biotin and streptavidin alkaline phosphatase conjugate. This assay gave comparable results for PLRV detection in potato to an assay based on immunoglobulins. The V3HCL-biotin preparations were stable and retained specific activity for more than 1 year when stored at 4 °C or -20 °C. The results demonstrate that scFv reagents derived from synthetic phage display platforms can provide effective alternatives to assays incorporating immune reagents. © 2009 Elsevier B.V. All rights reserved.

Publication metadata

Author(s): Al-Mrabeh A, Ziegler A, Cowan G, Torrance L

Publication type: Article

Publication status: Published

Journal: Journal of Virological Methods

Year: 2009

Volume: 159

Issue: 2

Pages: 200-205

ISSN (print): 0166-0934

ISSN (electronic): 1879-0984

Publisher: Elsevier BV


DOI: 10.1016/j.jviromet.2009.03.025

PubMed id: 19490974


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Funder referenceFunder name
Ministry of Higher Education of Syria
Scottish Government Rural Environment Research and Analysis Directorate