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

摘要

cqvip: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.