Preparation and epitope characterization of monoclonal antibodies suitable for detection of Tomato yellow leaf curl virus |
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Authors: | Leonardo J. Solmesky Avi Zrachya Galina Denisova Yedidya Gafni Jonathan M. Gershoni |
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Affiliation: | (1) Department of Cell Research and Immunology, Tel-Aviv University, Tel Aviv, 69978, Israel;(2) Department of Genetics, Agricultural Research Organization, The Volcani Center, Bet Dagan, 50250, Israel;(3) The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat Gan, 52900, Israel; |
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Abstract: | Tomato yellow leaf curl virus (TYLCV) is a begomovirus that seriously threatens tomato crops worldwide. Current immunodiagnostic methods for this pathogen employ commercially produced mAbs raised against TYLCV. However, despite the existence of these mAbs, little information regarding their characterization or strategy of production has been published. In addition, research on TYLCV would certainly benefit were more mAbs available, thus allowing efficient examination of the virus life cycle, modes of pathogenesis and possibly the development of resistant cultivars. The coat protein (CP) of TYLCV is the only known building block of the viral capsid. Therefore, in this study we used CP as an immunogen for the production of novel mAbs. We employed a strategy in which the CP was truncated at its C-terminus to avoid intra- and inter-protein interactions that could impair epitope exposure. For the same reason, we used a denaturated antigen to expose linear epitopes during the immunization. This effort yielded three mAbs: they were characterized biochemically and immunologically, and their epitopes were mapped. Possible applications of these mAbs are discussed. |
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