^(125)I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 ^(125)I

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。

A 18 mer oligonucleotides sequence is synthesized and grafted in 5′ position with a tyramine group. The tyramine derivatized oligonucleotides is labeled with ^(125)I using the chloramines-T method. The stability of ^(125)I-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 ^(125)I-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.