SEPARATION AND PURIFICATION OF BOVINE C-REACTIVE PROTEIN USING HYBRIDOMA PRODUCING MONOCLONAL ANTIBODY AGAINST BOVINE C-REACTIVE PROTEIN AND SAID ANTIBODY

摘要

PURPOSE:To separate and purify a C-reactive protein which is an antigen in high purity and yield, by preparing a hybridoma producing an antibody specifically reactive with a bovine C-reactive protein and using the antibody. CONSTITUTION:A mouse is immunized with a mixture of a bovine C-reactive protein (hereinafter abbreviated to CRP) or a fraction containing the CRP with Freund's complete adjuvant and splenic lymphocytes collected from the spleen of the mouse are used to mouse myelomatous cells by a well-known method to give a hybridoma producing antibovine CRP antibody. A supernatant of a culture medium after cultivating the hybridoma is salted out with ammonium sulfate to afford a precipitated protein, which is then purified with a protein A-Sepharose CL4B or aninon exchanger to provide a monoclonal antibovine CRP antibody. The resultant antibody is then linked and immobilized with a carrier for affinity chromatography and CRP is then separated from bovine blood serum using a column of the above-mentioned carrier.