VIEW ARTICLE

Techniques

Methods for the Enrichment of Desired B-Cell Populations Before Anti-Cauliflower Mosaic Virus Hybridoma Formation. R. A. George, Former graduate student, United States Department of Agriculture, Agricultural Research Service, Department of Botany and Plant Pathology, Oregon State University, Corvallis; R. H. Converse, Research plant pathologist, United States Department of Agriculture, Agricultural Research Service, Department of Botany and Plant Pathology, Oregon State University, Corvallis. Phytopathology 78:1631-1636. Accepted for publication 16 May 1988. This article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 1988. DOI: 10.1094/Phyto-78-1631.

Relative to a standard procedure and adjusted to uniform population sizes, immune complex masking, solid-phase immunoadsorption, and complement-mediated cytotoxicity used before murine hybridoma formation reduced by 49, 80, and 77%, respectively, the undesired anti-healthy Chinese cabbage (HCC) clones produced during studies on the production of anti-cauliflower mosaic virus (CaMV) monoclonal antibodies by hybridoma formation made from spleen cells from mice immunized with partially purified preparations of CaMV from infected Chinese cabbage leaves. These reductions led to 49, 72, and 54% increases, respectively, in the number of anti-CaMV clones found during screening. Analysis of the murine splenocyte populations by plaque-forming cell assay showed that these procedures resulted in reductions of anti-HCC B-cell populations of 49% for immune complex masking, 80% for solid-phase immunoadsorption, and 77% for complement-mediated cytotoxicity, compared to levels found by using a standard fusion procedure, thus enriching the level of desired anti-CaMV B cells in the resulting suspensions.