Cryopreservation of newly formed hybridomas

Abstract

A new freezing technique is described which permits time-consuming protocols as a first screening of newly formed hybridomas. In this procedure complete 96 well clustertrays with growing hybridomas are cryopreserved after programmed freezing. This procedure has been successfully applied to a number of fusion protocols for which the objective was to obtain monoclonal antibodies against tissue specific antigens. To this end hybridoma supernatants were screened by an immunoperoxidase technique on frozen sections. Freezing of the hybridoma containing clustertrays permitted extensive screening and partial characterization of the previously collected supernatants. After subsequent thawing of appropriate wells, the hybridoma clones proved to be viable and usually no loss of antibody production was observed.