A RAPID NONSELECTIVE METHOD TO GENERATE QUADROMAS BY MICROELECTROFUSION

A simple non-selective methodology was developed and standardized to g enerate desired hybrid-hybridoma or quadroma secreting bifunctional an tibodies. This novel protocol is based on microelectrofusion on a mean der chamber using a few hundred cells of each of the two parental hybr idomas with no laborious drug selection procedures. Seeding similar to 10 cells per well in a 96-well microtitre plate after fusion in 200 m u l standard medium containing 20% FBS and 10% Origen growth factor ge nerated positive quadromas secreting bispecific antibodies with good s tability after the second redone. Compared to the conventional PEG fus ion and other methods this simple protocol is both rapid and economica l. Generally, conventional methods to make quadromas and triomas requi re the introduction of drug selection markers into one or both of the parental cells, a procedure that could take 3-6 months. Utilizing the non-selective microelectrofusion method described here, we have genera ted several quadromas in a very short time. Further, such a protocol c ould also be potentially adopted to generate human hybridomas with few B cells isolated from peripheral blood lymphocytes enriched by antige n specific panning or affinity microelectrofusions.