Murine Monoclonal Antibodies Produced Against Two Illinois Strains of Barley Yellow Dwarf Virus: Production and Use for Virus Detection

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Abstract
A hybridoma cell line that secretes antibodies produced against an Illinois strain of barley yellow dwarf virus (BYDV) transmitted nonspecifically by Rhopalosiphum padi and Sitobion avenae (BYDV-PAV-IL) and two lines that secrete antibodies produced aginst an Illinois strain transmitted specifically by R. padi (BYDV-RPV-IL) were produced by somatic cell fusion between mouse myeloma cell line SP2/0-Ag14 and spleenocytes from BALB/c mice immunized with either virus strain. Ascitic fluid produced by clones PAV-IL-1, RPV-IL-1, and RPV-IL-5 had titers of 108, 108, and 109, respectively, in triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) with sap extracted from virus-infected plants. All three ascitic fluids had titers of 103 in ELISA with microtiter plates coated at pH 9.6 with purified virus for 2 hr at 37C. Immunoglobulin subclasses were IgG1 for PAV-IL-1 and RPV-IL-1 and IgG2a for RPV-IL-5, all with kappa light chains. Each of the antibodies was used sucessfully to trap homologous virus in double-antibody sandwich ELISA. In TAS-ELISA, clone PAV-IL-1 detected 17 of 17 PAV-like isolates in dried tissues from four U.S. states and 10 other countries. Clones RPV-IL-1 and RPV-IL-5 both detected seven of seven RPV-like isolates in dried tissue from three states and three outher countries. None of the clones detected any of 13 other BYDV isolates tested.