BINDING OPTIMIZATION OF AFP WITH ITS ANTIBODY BY IRMA METHOD IN TISSUE HOMOGENATE OF SOME OVARIAN TUMORS

Immunoradiometric assay (IRMA) was modified and used to characterize the binding of 125I-anti AFP antibody with its AFP antigen of benign germ cell (OI), malignant germ cell (OII), malignant serous (OIII) and isolated AFP from malignant germ cell ovarian tumor tissues. Different factor′s (concentrations of antibody, concentration of antigen, pH, time, temperature, and different concentrations of ions) affecting this binding were extensively studied. The observed optimum conditions for the binding were as follow: For (benign germ cell OI, malignant germ cell OII, malignant serous OIII and for isolated AFP of malignant germ cell), the optimum protein concentration in tissue homogenate was (300, 200, 200 and 75mg.ml-1), respectively. 125I-anti AFP antibody optimum concentration was (500, 400, 500 and 300mg.ml-1), respectively. Optimum pH was (8.0, 8.0, 7.6 and 8.0) respectively. The optimum time and temperature were 120 minute at 25˚C for (OI, OII, OIII) and180 minute at 4˚C for isolated AFP of malignant germ cell. The use of different divalent cations showed a decrease in the binding of AFP with its antibodies except for Mn+2 in OI, OII and OIII. In addition to a decreases the binding of AFP with its antibodies by using different halides in OI, OII and OIII.