Production of Monoclonal Antibodies Against Three Ilarviruses and Alfalfa Mosaic Virus and Their Use in Serotyping

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Abstract
Fourteen stable hybridoma cell lines that secrete monoclonal antibodies against prunus necrotic ringspot (NRSV), apple mosaic (ApMV), tobacco streak (TSV), or alfalfa mosaic (AMV) viruses were produced by fusing spleen cells of mice immunized with a mixture of all 4 viruses to mouse myeloma cell lines NS1/1 or P3 X63Ag8.653. Of the 7 hybridomas secreting monoclonal antibody specific for NRSV or ApMV, 2 were NRSV specific 3 were ApMV specific and 2 were cross-reactive for some strains of NRSV and ApMV. Five hybridomas secreted antibodies specific for TSV, and 2 hybridomas secreted antibody specific for AMV. High-titered ascitic fluid was produced to all of the hybridomas and used in indirect ELISA [enzyme-linked immunosorbent assay] to serotype a panel of isolates of NRSV, ApMV and TSV. Three serotypes of NRSV 5 serotypes of ApMV and 4 serotypes of TSV were identified. Titers of hybridoma ascitic fluid (measured by indirect ELISA) ranged from a low of 5000-62,500 for antibody. NA49F8 toward ApMV-F and NRSV-G, to a high of 7,812,000 for N63F10 toward NRSV-G. Only 5 of the 14 monoclonal antibodies precipitated homologous virus in agar double-diffusion assays. These monoclonal antibodies should prove to be valuable reagents for virus classification and disease diagnosis.