The use of in vitro immunisation, as an adjunct to monoclonal antibody production, may result in the production of hybridomas secreting polyreactive antibodies

The possibility that immortalisation of in vitro immunised splenocytes may result in hybridomas secreting polyreactive antibodies was investigated. A panel of nine murine hybridomas, secreting IgM(kappa) anti-goat immunoglobulin G (anti-GIgG), was produced by immortalising splenocytes that had been immunised in vitro with GIgG. The ability of the corresponding monoclonal antibodies (Mabs) to bind multiple antigens was investigated using two techniques. First, the affinity constants characterising the interactions of each of the nine Mabs with each of a panel of six antigens were determined. Second, the specific anti-GIgG activities of each hybridoma supernatant and its corresponding affinity-purified IgM fraction were determined and compared. In total, these experiments indicated that eight of the nine hybridomas were polyreactive.