VIEW ARTICLE

Techniques

Production of Monoclonal Antibodies Against Three Ilarviruses and Alfalfa Mosaic Virus and Their Use in Serotyping. Edward L. Halk, The American Type Culture Collection, Rockville, MD, Current address: Agrigenetics Advanced Research Laboratory, 5649 East Buckeye Road, Madison, WI 53716; H. T. Hsu(2), J. Aebig(3), and J. Franke(4). (2)(3)(4)The American Type Culture Collection, Rockville, MD. Phytopathology 74:367-372. Accepted for publication 6 October 1983. Copyright 1984 The American Phytopathological Society. DOI: 10.1094/Phyto-74-367.

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 four viruses to mouse myeloma cell lines NS1/1 or P3 X63Ag8.653. Of the seven hybridomas secreting monoclonal anitbody specific for NRSV or ApMV, two were NRSV specific, three were ApMV specific, and two were cross-reactive for some strains of NRSV and ApMV. Five hybridomas secreted antibodies specific for TSV, and two hybridomas secreted antibody specific for AMV. High-titered ascitic fluid was produced to all of the hybridomas and used in indirect ELISA to serotype a panel of isolates of NRSV, ApMV, and TSV. Three serotypes of NRSV, five serotypes of ApMV, and four serotypes of TSV were identified. Titers of hybridoma ascitic fluid (measured by indirect ELISA) ranged from a low of 5,000- 62,500 for antibody NA49F8 toward ApMV-F and NRSV-G, to a high of 7,812,000 for N63F10 toward NRSV-G. Only five 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.