Saevels J, Van Schepdael A, Hoogmartens J
Laboratory for Pharmaceutical Chemistry and Drug Analysis,K.U Leuven, Belgium.
J Capillary Electrophor. 1997 Jul-Aug;4(4):167-72.
A capillary electrophoresis system that integrated an enzymatic reaction and capillary polymer sieving electrophoresis was used to check the enzymatic stability of oligonucleotides. Phosphodiesterase I was employed to assess the susceptibility to 3'-exonucleolytic breakdown of some unmodified and modified oligonucleotides. Before degradation, the purity of the synthetic oligodeoxynucleotides was checked by capillary electrophoresis with a replaceable hydroxyethyl cellulose polymer solution. Enzymatic breakdown was achieved inside the capillary by merging substrate and enzyme zones based on their difference in electrophoretic mobility. After reaction, oligonucleotide fragments were swept to the detector, where they were individually detected and the remaining substrate was quantified. The results from the in-capillary degradation were compared to an off-line incubation and separation.
使用集成了酶促反应和毛细管聚合物筛分电泳的毛细管电泳系统来检测寡核苷酸的酶稳定性。采用磷酸二酯酶I评估一些未修饰和修饰寡核苷酸对3'-核酸外切酶降解的敏感性。在降解之前，通过使用可替换的羟乙基纤维素聚合物溶液的毛细管电泳检查合成寡脱氧核苷酸的纯度。基于底物和酶区在电泳迁移率上的差异，通过合并底物和酶区在毛细管内实现酶促降解。反应后，寡核苷酸片段被扫至检测器，在那里它们被单独检测，剩余底物被定量。将毛细管内降解的结果与离线孵育和分离的结果进行比较。
