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125I标记免疫球蛋白重链V1家族框架区反义寡核苷酸

Study on Labeling of Antisense Oligonucleotides of IgH V1 FR mRNA With 125I

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【作者】 王荣福邱峰张春丽

【Author】 WANG Rong-fu, QIU Feng, ZHANG Chun-li (Department of Nuclear Medicine, Peking University First Hospital, Beijing 100034, China)

【机构】 北京大学第一医院核医学科北京大学第一医院核医学科 北京 100034北京 100034北京 100034

【摘要】 用DNA合成仪将含酪胺的单体连接在18聚体寡核苷酸的5′端,并用氯胺-T法进行125I标记。该标记方法的标记率为80.7%,标记物放化纯度为98.7%,体外放置24h放化纯度仍高于90%。聚丙烯酰胺凝胶电泳(PAGE)显示各时间点的条带与未保温的对照组平行,均未见异常条带。

【Abstract】 A 18 mer oligonucleotides sequence is synthesized and grafted in 5′ position with a tyramine group. The tyramine derivatized oligonucleotides is labeled with 125I using the chloramines-T method. The stability of 125I-Tyramine-FR-ASON in blood is measured. The labeling yield, radiochemical purity and specific activity are 80.7%, 98.7% and 100 GBq/g, respectively. The radiochemical purity of 125I-Tyramine-FR-ASON is over 90% at 24 h in ambient atmosphere. The polyacrylamide gel electrophoresis results show that there is not any additional band at 2, 6 and 24 h post incubation. The radioiodination method by conjugation of the oligonucleotides with tyramine is very successful.

【关键词】 反义寡核苷酸125I标记
【Key words】 antisense oligonucleotides125Ilabeling
【基金】 国家自然科学基金资助项目(30270410)
  • 【分类号】R817
  • 【被引频次】8
  • 【下载频次】56
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