CAJ | 학술논문

目的测定一种新型~(99m)Tc定点标记膜联蛋白V与磷脂酰丝氨酸(PS)外翻红细胞的结合亲和力.方法通过毕赤酵母的培养和甲醇诱导表达获得带有金属螯合位点的膜联蛋白V;用超滤的方法纯化膜联蛋白V;用葡庚糖酸钠和氯化亚锡还原将~(99m)Tc定点标记于膜联蛋白V氨基末端.采用不同浓度钙离子滴定放射性膜联蛋白V结合磷脂酰丝氨酸外翻红细胞的亲和力.结果在不同的蛋门质分子/细胞比值下测定出的半数标记蛋白结合细胞的钙离子浓度是不同的,在低蛋白质分子/细胞比值下测定~(99m) Tc-膜联蛋白V对细胞的亲和力pK=33.4.结论毕赤酵母系统重组表达的膜联蛋白V具有较高的结合外露PS红细胞的能力.
목적 측정일충신형~(99m)Tc정점표기막련단백V여린지선사안산(PS)외번홍세포적결합친화력.방법 통과필적효모적배양화갑순유도표체획득대유금속오합위점적막련단백V;용초려적방법순화막련단백V;용포경당산납화록화아석환원장~(99m)Tc정점표기우막련단백V안기말단.채용불동농도개리자적정방사성막련단백V결합린지선사안산외번홍세포적친화력.결과 재불동적단문질분자/세포비치하측정출적반수표기단백결합세포적개리자농도시불동적,재저단백질분자/세포비치하측정~(99m) Tc-막련단백V대세포적친화력pK=33.4.결론 필적효모계통중조표체적막련단백V구유교고적결합외로PS홍세포적능력.
Objective To derermine the affinity of site-specific labeled annexin V with ~(99m)Tc to phosphatidylserine (PS)exposed erythrocytes.Methods The annexin V fused with a metal chelating binding site was obtained from Pichia Pastoris culture and methanol induction expression.The annexin V was purified from the culture supernatant crude product by uhrafihration.The annexin V was conjugated with ~(99m)Tc site-specifically through sodium glucoheptonic acid and SnCl_2.The radioactive annexin V was added to determine its affinity to PS exposed erythrocytes.Results The calcium concentration at which half of the protein is bound to cells(EC50)is changed with varying ratio of protein to cells.The affinity was determined at a low protein/cell ratio as 33.4.Conclusion The annexin V recombinantly expressed in Pichia Pastoris shows high affinity to PS exposed erythrocytes.