Chemiluminescence Immunoassay for Measurement of Estrone-3-Sulfate Using Monoclonal Antibody to Estrone-3-Glucuronide

Estrone-3-Glucuronide에 대한 단일클론항체를 이용한 Estrone-3-Sulfate 측정을 위한 화학발광면역분석법

This study was carried out to develop an immunoassay for the diagnosis of the pregnancy and ovarian function of domestic animals. Using 2E92C10 monoclonal antibody(McAb) generated against estrone-3-glucuronide(E1-3-G) and appeared a high cross-reactivity with estrone-3-sulfate(E1-3-S), chemiluminesence immunoassay (CIA) to detect E1-3-S was developed. 2E92C10 McAb cross-reacted with E1-3-S (30%) was purified from ascites fluid using protein G sepharose gel column. The purity of purified antibody fraction was monitored by SDS-PAGE and was better compared to that of crude ascite fluid. The soild and liquid phase CIA for E1-3-S were established utilizing 2E92C10 antibody and E1-3-G-ABEI conjugate used as a tracer. As the results, the titer of 2E92C10 antibody was 5g/ml in soild phase and 1:2000 in liquid phase. The sensitivity on soild and solid phase CIA were about 200 pg/ml. These results indicate that CIA for measurement of E1-3-S was successfully developed by using ant-E1-3-G McAb cross-reacted with E1-3-S and could be usefully used to research this area.