1987 Fiscal Year Final Research Report Summary
IMPROVEMENT OF CELL FUSION METHOD
| Project/Area Number |
60870021
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
TACHIBANA Takehiko RESEARCH INSTITUTE FOR TUBERCULOSIS AND CANCER TOHOKU UNIVERSITY, 抗酸菌病研究所, 教授 (50076973)
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| Co-Investigator(Kenkyū-buntansha) |
KUDO Toshio COLLEGE OF MEDICAL SCIENCES TOHOKU UNIVERSITY, 助教授 (70091684)
TADA Masao RESEARCH INSTITUTE FOR TUBERCULOSIS AND CANCER TOHOKU UNIVERSITY, 抗酸菌病研究所, 助教授 (10006083)
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| Project Period (FY) |
1985 – 1987
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| Keywords | CELL FUSION / MOUSE MONOCLONAL ANTIBODY / HUMAN MONOCLONAL ANTIBODY / GIT培地 |
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| Research Abstract |
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.
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[Publications] SUZUKI, N., KUDO, T., SHIRAKO, Y., EHARA, Y., AND TACHIBANA, T.: "PREPARATION OF MONOCLONAL ANTIBODIES TO CYLINDRICAL INCLUSION, AMORPHOUS INCLUSION, AND CAPSID PROTEINS OF WATERMELON MOSAIC VIRUS 2 AND ANALYSIS OF THEIR REACTIVITIES." VIROLOGY. -IN PREPARATION
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