IMPROVEMENT OF CELL FUSION METHOD

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1. THE FOLLWING TECHINQUES WERE ATTEMPTED TO DEVELOP GENERAL SELECTION METHOD FOR HYBRID CELLS; (1) SORTING OF FUSION CELLS WITH TWO FLUORESCENT COLORS BY FLOW CYTOMETRY AFTER CELL FUSION OF TWO KINDS OF PARENTAL CELLS LABELED WITH FITC- AND XRITC-OCTADECYLAMINE RESPECTIVELY. (2) HYBRID CELL SELECTION BY USING ONE OF PARENT CELLS WHICH PROTEIN SYNTEHSIS WAS BLOCKED WITH ACTINOMYCIN D AND EMETINE HYDROCHLORIDE. (3) CELL FUSION AFTER CONJUGATION WITH AVIDIN OF BOTH BIOTINYLED MYELOMA CELLS AND B LYMPHOCYTES REACTING WITH BIOTINYLED ANTIGEN. MANY EFFORTS WERE DONE BUT FAILED TO PRODUCE ANY NEW EFFECIENT SELECTION METHOD.2. IT WAS FOUND THAT A NEW GIT CULTURE MEDIUM IS MOST SUITABLE FOR EFFECIENT PRODUCTION OF B CELL HYBRIODOMA; (1) THE FUSION EFFICIENCY OF MOUSE B CELL HYBRIDOMA WAS 1/8,000 MYELOMA CELLS. THIS IS THE BEST EFFICIENCY OVER THOSE REPORTED SO FAR. (2) EFFECIENT PRODUCTION OF HUMAN MONOCLONAL ANTIBODY WAS ALSO OBTAINED BY CELL FUSION BETWEEN (MOUSX X HUMAN) HETEROMYELOMA CELLS (SHM-D33) AND EBV-TRANSFORMED B CELLS AND SELECTION OF HYBRID CELLS IN GIT MEDIUM. COMPARISONS OF FUSION AND CLONING EFFICIENCY IN BETWEEN GIT AND ORDINARY CULTURE MEDIUM WERE 1/5,500 VS. 1/11,000 AND 43.3-56.7% VS. 3.3-13.3% RESPECTIVELY.