Mouse-Anti-Muc4放射性碘标记方法探索及质量鉴定

发布时间：2018-05-05 21:28

本文选题：~(125)I + 分子探针　；参考：《川北医学院》2012年硕士论文

【摘要】：目的：探索125I-Mouse-Anti-Muc4标记方法，选择最佳标记条件，并进行质量鉴定，为进一步了解分子探针在体内的生物学行为以及胰腺癌的放射免疫显像和治疗奠定基础。方法：本实验分别采用Iodogen、NBS作为氧化剂制备分子探针—125I-Mouse-Anti-Muc4，其它条件固定时，依次改变反应温度、时间和碘与抗体、Iodogen/ NBS与抗体用量，用纸层析法测定标记产物的标记率，探索最佳的标记反应条件；然后将反应物经PD-10脱盐柱淋洗，收集蛋白峰管，测定放射性活度，并测定纯化后的放化纯；并测定标记物在不同储存溶剂中的体外稳定性。结果：Iodogen、NBS法标记Mouse-Anti-Muc4的最高标记率分别为（81.3±0.97）%、（50.7±0.79）%，Iodogen为比较理想的氧化剂；不同的反应温度、时间和碘与抗体、Iodogen/ NBS与抗体用量可影响标记效果，当在室温（25~0C）下反应15min，，碘/抗体比值为1.5，Iodogen/抗体比值为10时标记率最高；~(125)I- Mouse-anti-Muc4的放化纯达（95.6±1.26）%；125I-Mouse-anti-Muc4在生理盐水及1%胎牛血清中放置48小时内放化纯维持在90%左右。结论：首次使用~(125)I标记Mouse-anti-Muc4成功获得了分子探针—125I-Mouse-anti-Muc4；Iodogen法标记Mouse-anti-Muc4是比较理想的方法，能够得到比较理想的标记率及纯化率；不同的反应温度、时间和碘与抗体、Iodogen/ NBS与抗体用量可影响其标记效果；~(125)I-Mouse-anti-Muc4体外稳定性好；分子探针—~(125)I-Mouse-anti-Muc4的免疫活性、生物学行为及其胰腺癌的放射免疫显像和治疗等方面值得进一步研究。
[Abstract]:Objective: to explore the method of 125I-Mouse-Anti-Muc4 labeling, to select the best labeling conditions, and to identify the quality of the probe, so as to lay a foundation for further understanding the biological behavior of the molecular probe in vivo and the radioimmunoimaging and treatment of pancreatic cancer. Methods: the molecular probe -125I-Mouse-Anti-Muc4 was prepared by using Iodogen-NBS as oxidant. When the other conditions were fixed, the reaction temperature, reaction time and the amount of iodide / NBS and antibody were changed in turn. The labeling rate of labeled products was determined by paper chromatography. The best labeling reaction conditions were explored, then the reactants were eluted by PD-10 desalination column, protein peak tubes were collected, radioactivity was determined, radiochemical purity was determined, and the stability of the reactants in different storage solvents in vitro was determined. Results the highest labeling rates of Mouse-Anti-Muc4 labeled by the 7% Iodogen method were 81.3 卤0.97 and 50.7 卤0.79, respectively, and Iodogen was the ideal oxidant. Different reaction temperature, reaction time, iodine / NBS and antibody dosage could affect the labeling effect. The radiochemical purity of 125I-Mouse-anti-Muc4 was 95.6 卤1.26C125I-Mouse-anti-Muc4 in normal saline and 1% fetal bovine serum for about 90% within 48 hours after the reaction at room temperature for 15 min, and the ratio of iodine / antibody was 1.5% Iodogen-antibody / antibody ratio was 10:00. The radiochemical purity of 125I-Mouse-anti-Muc4 in normal saline and 1% fetal bovine serum was about 90%. Conclusion: the molecular probe -125I-Mouse-anti-Muc4- Iodogen method was successfully obtained for the first time in the labeling of Mouse-anti-Muc4, and the labeling rate and purification rate were obtained, and the reaction temperature was different, and the molecular probe -125I-Mouse-anti-Muc4- Iodogen method was used to label Mouse-anti-Muc4 successfully for the first time. Time and dosage of Iodogen / NBS and antibody could affect the stability of 125I-Mouse-anti-Muc4 in vitro, and the immunological activity, biological behavior and radioimmunoimaging and treatment of pancreatic cancer could be further studied by molecular probe -125I-Mouse-anti-Muc4.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R817.4;R735.9

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