| 钙滴定法测定~(99m)Tc定点标记膜联蛋白V的亲和力 |
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| 引用本文： | 蔡炯,李方,牛娜,王世真.钙滴定法测定~(99m)Tc定点标记膜联蛋白V的亲和力[J].基础医学与临床,2010,30(5):534-537. |
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| 作者姓名： | 蔡炯 李方 牛娜 王世真 |
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| 作者单位： | 中国医学科学院,北京协和医院核医学科,北京,100730 |
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| 基金项目： | 北京协和医院临床重点课题;;霍英东肺癌专项基金 |
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| 摘 要： | 目的测定一种新型99mTc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99mTc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。结果在不同的蛋白质分子/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/细胞比值下测定出99mTc-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。
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| 关 键 词： | 膜联蛋白V 亲和力 99mTc 毕赤酵母 磷脂酰丝氨酸 |
| 收稿时间： | 2009-9-1 |
| 修稿时间： | 2009-11-24 |
The measurement of affinity of site-specific labeled annexin V with ~(99m)Tc by calcium titration |
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| CAI Jiong,LI Fang,NIU Na,WANG Shi-zhen.The measurement of affinity of site-specific labeled annexin V with ~(99m)Tc by calcium titration[J].Basic Medical Sciences and Clinics,2010,30(5):534-537. |
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| Authors: | CAI Jiong LI Fang NIU Na WANG Shi-zhen |
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| Institution: | Department of Nuclear Medicine;PUMC Hospital;Chinese Academy of Medical Sciences;Beijing 100730;China |
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| Abstract: | Objective To derermine the affinity of site-specific labeled annexin V with 99mTc 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 ultrafiltration.The annexin V was conjugated with 99mTc site-specifically through sodium glucoheptonic acid and SnCl2.The radioactive annexin V was added to determine its... |
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| Keywords: | annexin V affinity 99mTc Pichia Pastoris phosphatidylserine |
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