125I标记氧化低密度脂蛋白抗体

Labeling of OxLDL-Ab With 125I

  • 摘要: 采用Iodogen法对氧化低密度脂蛋白抗体(OxLDL-Ab)进行了125I标记,并对标记条件进行了优化;采用PD-10柱对标记抗体进行分离纯化,并对125I-OxLDL-Ab的体外稳定性和活性进行评价。结果表明,125I-OxLDL-Ab标记率>80%,放化纯度>95%,比活度达73.3 TBq/g;125I-OxLDL-Ab分别在生理盐水、含2%BSA的生理盐水和人血清中4 ℃放置96 h,放化纯度降低均小于5%;125I-OxLDL-Ab在体外与OxLDL结合的Ka为10.6±1.3 GL/mol。以上结果提示125I-OxLDL-Ab体外稳定性好,与OxLDL结合活性高,125I标记对其活性影响小,可以用于体外放免诊断试剂盒的研究。

     

    Abstract: OxLDL-Ab was labeled with 125I by the Iodogen method, and the labeling was purified by PD-10 column. The optimal condition of radiolabeling reaction was determined. The stability and affinity to OxLDL of 125I-OxLDL-Ab in vitro were evaluated as well. The labeling yield of 125I-OxLDL-Ab was more than 80% and the radiochemical purity was more than 95%. The results of in vitro stability studies showed that the radiochemical purity of 125I-OxLDL-Ab declined less than 5% after 96 h storage under following condition: 0.9%NaCl, 2%BSA in 0.9%NaCl solution and human serum at 4 ℃ respectively. In the binding experiments of 125I-OxLDL-Ab and OxLDL, Ka was (1.06±0.13)×1010 L/mol. The results showed that 125I-OxLDL-Ab had high stability and high affinity to OxLDL, so it was worthy of further research for in vitro diagnosis kit.

     

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