Selection of specific single chain variable fragments (scFv) against Polymyxa betae from phage display libraries
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Department of Plant Viruses, Iranian Research Institute of Plant Protection, P.O. Box 1452-19395, Tehran, Iran
Nanosystems Research Team (NRT), Department of Microbial Biotechnology and Biosafety, Agricultural Biotechnology Research Institute of Iran (ABRII), P.O. Box 31535-1897, Karaj, Iran
Department of Agricultural Biotechnology, Shahed University, P.O. Box 18155-159, Tehran, Iran
Department of Plant Pathology, Tarbiat Modares University, P.O. Box 336-14115, Tehran, Iran
Department of Plant Pathology, Sugarebeet Seed Research Institute, P.O. Box 4114-31585, Karaj, Iran
Mohammad Reza Safarnejad
Department of Plant Viruses, Iranian Research Institute of Plant Protection, P.O. Box 1452-19395, Tehran, Iran
Submission date: 2013-09-03
Acceptance date: 2013-10-04
Journal of Plant Protection Research 2013;53(4):357–363
Sugar beet is one of the most important industrial crops in Iran. For the last two decades it has been mainly affected by a destructive virus, beet necrotic yellow vein virus (BNYVV). The Polymyxa betae is the only natural transmitting agent of the disease among the plants. Developing accurate diagnostic methods may have a major impact on the rising of resistant germplasms. In the present study, specific monoclonal recombinant antibodies in the form of single chain variable fragments (scFv) were obtained from naïve phage display libraries. The fungus specific glutathione-S-transferase (GST) protein was chosen as an antigen for developing antibodies and diagnostic purposes. To generate specific scFv, screening of Tomlinson phage display libraries was performed by applying both recombinant and native fungal GST. Using the recombinant GST in the panning process resulted in the isolation of an antibody only bound to recombinant GST but it failed to detect native GST in the infected plants. Alternatively, the process of panning was carried out by applying native fungal GST trapped to immunotubes through specific polyclonal antibody intermediate. The recent approach resulted in the selection of a specific scFv binding to native GST which was able to detect the presence of the fungi within infected plants. To the best of our knowledge, this is the first report on the generation of recombinant antibodies against Polymyxa betae, fungal vector of sugar beet rhizomania disease.
The authors have declared that no conflict of interests exist.
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