Monoclonal Antibodies Against the Whitefly-Transmitted Tomato Yellow Leaf Curl Virus and Their Application in Virus Detection

Tomato yellow leaf curl virus（TYLCV）is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein（CP）gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21（DE3）using pET-32a as the expression vector.The recombinant protein was purified through Ni＋-NTA affinity column and used to immunize BALB/c mice.Three hybridoma cell lines（2B2,2E3 and 3E10）secreting monoclonal antibodies（MAbs）against TYLCV CP were obtained by fusing mouse myeloma cells（Sp 2/0）with spleen cells from the immunized BALB/c mouse.The titers of ascitic fluids of three MAbs ranged from 10-6 to 10-7 in indirect-ELISA.Isotypes and subclasses of all the MAbs belonged to IgG1,κ light chain.Triple antibody sandwich enzyme-linked immunosorbent assay（TAS-ELISA）showed that the MAb 3E10 could react with five begomoviruses infecting tomato,while the other two（2B2 and 2E3）mainly reacted with TYLCV.TAS-ELISA was set up using the MAb 3E10,and the established method could successfully detect virus in plant sap at 1：2 560（w/v,g mL-1）.Detection of field samples showed that begomoviruses were common in tomato crops in Zhejiang Province,China.