Lebedev Alexandre V, Combs David, Hogrefe Richard I
Trilink Biotechnologies, Inc., San Diego, California 92121, USA.
Bioconjug Chem. 2007 Sep-Oct;18(5):1530-6. doi: 10.1021/bc0603891.
A C10 linker phosphoramidite reagent terminated with a succinimidyl-activated carboxyl group was prepared and used to couple to the 5'-end of an oligonucleotide synthesized on a solid support. The succinimidyl-activated carboxyl functionality can be used for rapid conjugation of amines to oligonucleotides on solid support or it can be hydrolyzed to form a carboxylic acid functionality. The activated linker was successfully used for conjugation of several primary and secondary aliphatic amine derivatives (including biotin and fluorescein cadaverine) onto a solid support-bound 12-mer DNA oligonucleotide at scales ranging from 0.15 to 1.0 micromol. The overall yields of the conjugation products after AMA deprotection and cleavage from the solid support ranged from 43 to 75% of the total oligonucleotide product. This value is significant, as it includes oligonucleotide synthesis, coupling of the linker, and conjugation of the amine. In addition, the entire process of oligonucleotide synthesis, linker coupling, amine conjugation, deprotection, and cleavage of the oligonucleotide from solid support can be accomplished in 1 day.
制备了一种以琥珀酰亚胺基活化羧基终止的C10连接亚磷酰胺试剂，并将其用于与在固相载体上合成的寡核苷酸的5'-末端偶联。琥珀酰亚胺基活化的羧基官能团可用于将胺快速偶联到固相载体上的寡核苷酸上，或者它可以水解形成羧酸官能团。该活化连接子成功用于将几种伯胺和仲胺衍生物（包括生物素和荧光素尸胺）以0.15至1.0微摩尔的规模偶联到固相载体结合的12聚体DNA寡核苷酸上。在AMA脱保护并从固相载体上切割后，偶联产物的总产率为总寡核苷酸产物的43%至75%。这个值很可观，因为它包括寡核苷酸合成、连接子偶联和胺的偶联。此外，寡核苷酸合成、连接子偶联、胺偶联、脱保护以及从固相载体上切割寡核苷酸的整个过程可以在一天内完成。
