Purification of igg-monoclonal antibody and its use

PURPOSE: To obtain the subject highly purified antibody for use as immunotherapeutic agent, immunodiagnostic agent and the like by chromatographically separating a previously dialyzed solution containing an antibody on an anionic exchanger, a cationic exchanger and a hydrophobic phase followed by elution at the gradual gradient and sterilization. CONSTITUTION: A hybridoma cell which produces an antibody is proliferated as an ascites tumor in vivo or in a nutrient medium in vitro to give an antibody- containing solution, which is dialyzed by a buffer solution to give a starting solution containing the antibody, which is separated by the anionic exchanger chromatography and sterilized. Subsequently it is subjected to successive chromatographic separation by a cationic exchanger and by a hydrophobic phase, whereby the monoclonal antibody is bound to each chromatographic phase. The monoclonal antibody is washed, eluted at the gradual gradient and additionally subjected to a particular sterilization step using 0.01-1% tri-n-butyl phosphate or the like to give the monoclonal antibody having a purity of 99.5% or more for use as an intravenous application, an immunodiagnostic agent or the like in the immunotherapy. COPYRIGHT: (C)1995,JPO