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KMID : 0900919960200020215
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Korean journal of Animal Reproduction
1996 Volume.20 No. 2 p.215 ~ p.221
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Chemiluminescence Immunoassay for Measurement of Estrone-3-Sulfate Using Monoclonal Antibody to Estrone-3-Glucuronide
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Kim Yoon-Gyu
Kim Choon-Won
Kim Jong-Bae
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Abstract
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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.
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KEYWORD
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Monoclonal antibody, estrone-3-sulfate, chemiluminesence immunoassay
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