A NEW METHOD TO GENERATE QUADROMAS BY ELECTROFUSION AND FACS SORTING

Bispecific monoclonal antibodies (bsMAbs) are unique molecules incorpo rating two different paratopes in a single antibody molecule. BsMAbs c an be useful in different areas of research as well in clinical applic ations. Traditionally, bsMAbs are produced by hybrid-hybridomas that a re generated by the fusion of two pre-established hybridomas, The deve lopment of such hybrid-hybridomas can be difficult and time-consuming. Here, we introduce a new technique to generate such hybrids, electro- FACS-fusion, In this procedure, before the electrofusion, one of the h ybridomas is labeled with fluorescein isothiocyanate (FITC) and the ot her with tetramethylrhodamine isothiocyanate (TRITC). The mixture of c ells is then electrofused, and cells exhibiting dual fluorescence are selected by fluorescence activated cell sorting (FACS), The fused cell s are directly plated in microplates for clonal growth, Using this tec hnique, we produced three new hybrid-hybridomas secreting bsMAb that c ould be used for the next generation of immunoassays.