铁蛋白受体放射配体结合分析法 |
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引用本文: | 刘玉峰,李丰益,廖清奎,罗春华. 铁蛋白受体放射配体结合分析法[J]. 中国小儿血液与肿瘤杂志, 1997, 0(2) |
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作者姓名: | 刘玉峰 李丰益 廖清奎 罗春华 |
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作者单位: | 华西医科大学附二院小儿血液/肿瘤研究室!450052 |
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摘 要: | 本文采用差速离心法制备胎盘微绒毛膜,用125I-Hunter试剂联接标记铁蛋白,在国内首次建立了胎盘微绒毛膜铁蛋白受体放射配体分析法,并进行了最适反应条件及铁蛋白受体的特性研究。所标记的125I一铁蛋白放化纯达95%以上,且免疫活性和生物活性均较好。用放射配体受体分析测定了11例正常足月孕妇胎盘微绒毛膜铁蛋白受体数为(3.534土1.105)×1012位点/毫克膜蛋白,亲和力常数Ka为(2.203土0.622)×107mol-1.L。实验条件为:膜蛋白160ug,125I互铁蛋白与游离铁蛋白分离的PEG浓度为12%。对铁蛋白受体特性研究表明:铁蛋白受体与铁蛋白结合具有高度特异性、可饱和性和中度亲和力。
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关 键 词: | 胎盘微绒毛膜 铁蛋白受体 受体放射配体结合分析 |
THE RADIOLI GAND ASSAY ON HUMAN PLACENTAL FERRITIN RECEPTOR |
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Abstract: | Using the human placental microvillous membrane as the receptor reagent and Na125Iapo--Fr as the radioligand , the receptor radioligand asay (RRA) of ferritin receptor (FrR)was first established in our country. The microvillous membrane was isolated by the differential centrifugation, Na 125I--apo--Fr was labelled by modified 125I--Hunter conjugation labelling method. the experimental condition and the characteristic of FrR were also studied.The radiochemical purity of 125I--apo--Fr was above 95% and immunologic competence andbiological activity of 125I--apo --Fr were better. The analysis of FrR expression on placentswas one in 11 cases of normal term pregnant women. The result indicated that the FrR expression was (3. 534± 1. 105) X 1012, sites/mg. pro; Ka: (2. 203±0. 622) X 107 mol-1. L.the experimental condition were as following: the memberane protein 160mg, 125I--apo--Fr 0.lug, the receptor--bound ferritin and free ferritin were separated by polyethy--lenegly 12%The study of FrR characteristic revealed that the FrR binding Fr is of very high specificity,saturation, and moderate affinity. |
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