Habus I, Xie J, Iyer R P, Zhou W Q, Shen L X, Agrawal S
Hybridon, Inc., Cambridge, Massachusetts 02139, USA.
Bioconjug Chem. 1998 Mar-Apr;9(2):283-91. doi: 10.1021/bc970132q.
Conjugates of oligodeoxyribonucleotide phosphorothioate (ODN-PS) with folic acid, retinoic acid, arachidonic acid, and methoxypoly(ethylene glycol)propionic acid have been synthesized. The procedure involved the initial solid-phase preparation of 5'-amino-functionalized ODN-PS using N-pent-4-enoyl-derived (PNT) nucleoside phosphoramidites followed by conjugation of the oligonucleotide either to the ligand acids, using 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide as a coupling reagent, or to their corresponding succinimidyl derivatives. Subsequent exposure of the support to aqueous ammonium hydroxide (28%, 2 h, 55 degrees C) resulted in the release of the fully deprotected ODN conjugates, which were purified by reversed-phase HPLC or by preparative polyacrylamide gel electrophoresis. The identity of the oligonucleotide conjugates was confirmed by MALDI-TOF mass spectral analysis.
已合成了硫代磷酸寡脱氧核糖核苷酸（ODN-PS）与叶酸、视黄酸、花生四烯酸和甲氧基聚（乙二醇）丙酸的缀合物。该过程包括使用N-戊-4-烯酰基衍生的（PNT）核苷亚磷酰胺对5'-氨基官能化的ODN-PS进行初始固相制备，然后使用1-[3-（二甲基氨基）丙基]-3-乙基碳二亚胺作为偶联剂将寡核苷酸与配体酸或其相应的琥珀酰亚胺衍生物偶联。随后将载体暴露于氢氧化铵水溶液（28%，2小时，55℃），导致完全脱保护的ODN缀合物释放，通过反相高效液相色谱或制备型聚丙烯酰胺凝胶电泳对其进行纯化。通过基质辅助激光解吸电离飞行时间质谱分析确认了寡核苷酸缀合物的身份。
