RENIN MONOCLONAL ANTIBODY AND IMMUNOLOGICAL MEASURING METHOD FOR ACTIVE TYPE RESIN USING THE SAME

摘要

PURPOSE: To obtain a renin monoclonal antibody useful for diagnosing and treating hypertensive diseases, capable of measuring active type renin in high accuracy and sensitivity, produced by a hybridoma prepared by using a recombinant human renin. CONSTITUTION: A recombinant human renin and a Freund's perfect adjuvant are administered to a Balb/c mouse at intervals of three weeks and sensitized. After three days from the final immune, the spleen is collected and cleaned to give a splenocyte. Then the splenocyte is suspended in an isotonic mannitol solution and mixed with a mouse myeloma cell. The prepared mixed solution is put in a parallel chamber of an electrically fusing device, a DC pulse is impressed at 420V to the mixed solution to fuse both the cells and the fused cell is selectively cultured in a HAT medium to give a hybridoma cell. Then the cell is cloned into a monoclone by a limiting dilution and cultured to give the objective renin monoclonal antibody inhibiting >=50% renin enzyme activity at 5×10<-9> M antibody concentration.