We have established an Omni-Hybridoma™ platform, in which newly fused hybridoma cells are subcloned, grown and selected using an agarose-based semi-solid medium containing growth factors, B-cell stimulators and medium supplements optimized for the growth of single hybridoma cells. This revolutionary platform ensures that a large number of hybridoma clones can be selected after each cell fusion.
In traditional hybridoma plating methods, multiple rounds of limiting dilutions are required to obtain monoclonal hybridoma clones. A serious problem is that some hybridoma clones overgrow others prior to cloning. Often, these faster growing cell lines do not synthesize antibodies, resulting in a failure to obtain the best antibody-producing hybridoma clones. Also, it is extremely labor-intensive and time-consuming to single-cell clone a hybridoma clone by performing multiple rounds of limited dilutions, thus making it difficult to isolate a large number of hybridoma clones from each cell fusion.
The Omni-Hybridoma™ platform is designed to select and clone hybridoma cell clones immediately after fusion. This eliminates the possibility of overgrowth of potentially valuable slow-growing clones by fast-growing clones. In this platform, hybridoma colonies are clonal from the start; the number of clones to be screened for the secretion of a specific antibody is minimized because all daughter cells are found in the same colony. Most importantly, since HAT selection and cloning of hybridomas are performed simultaneously in a single step, large numbers of hybridomas can be selected and tested. In fact, more than 1,500 hybridoma clones can be single-cells cloned in ten 15cm dishes after a single fusion, thus significantly increasing the number of candidate hybridoma clones that are specific for the target immunogen.