Optimization For Preparation of Ca-125 Irma-Kit

Optimization on the preparation of Ca-125 IRMA-kit has been studied involving preparation of 125I-labeled monoclonal anti Ca-125 as a tracer and preparation of coated tubes. Chloramine-T and N-bromosuccinimide were used as oxidators by varying them in their quantity. optimizations on preparation of coated tube was carried out by variation in volume of coating solution, effect of washing, addition of BSA 0.1%, buffer and pairing test for the monoclonal anti CA-125. The use of chloramine-T as oxidator for 125I-labeling of monoclonal anti Ca-15 was found to give labeling-yield of about 96.5% and radiochemical purity of 93.2%. Optimum amount of the oxidizing agent was found to be 10 μg. There were not any significant differences in utilization of coating solution volume ranging at 300 to 600 μL for preparing monoclonal anti Ca-125 coated tube. The non specific binding (NSB) and the %B/T values were found to be unaffected by washing step, while a blocking of coated tube using of BSA 0.1 % decreased the %B/T value. Utilization of bicarbonate buffer for the preparation of coated tube was found to be more satisfactory than that of phosphate buffer. It was shown that the monoclonal anti Ca-125 M37203M used for tracer being paired with monoclonal anti Ca-125 M86924M used for coating solution gave an optimal result based on the values of %NSB and %B/T