硫代反义寡聚核苷酸在线性聚丙烯酰胺-毛细管电泳中的分离行为

通过对硫代反义寡聚核苷酸在毛细管线性聚丙烯酰胺胶（LPA）中的电泳行为的研究，发现在10％浓度的LPA和100mmol/L Tris-borate及7mol/L尿素的缓冲溶液中（pH=8.2），这类被分析物质有着较好的分离效果和重现性，能使相差一个碱基的硫代反义寡聚核苷酸片断得到基线分离。非常适合用于对合成的反义寡聚核苷酸的定性分析并进一步应用于纯度测定。

Separation Investigation of Antisense Phosphorothioate Oligonucleotides Using Capillary Electrophoresis with Linear Polyacrylamide

The capillary electrophoresis (CE) with linear polyacrylamide (LPA) was employed to separate the phosphorothioate oligonucleotides (PS-ODNs). The content of 10% linear polyacrylamide was used as a replaceable sieving matrix and 100 mmol/L Tris-borate buffer containing 7 mol/L urea at pH 8.2 was chosen as running buffer. The running voltage was designed at 330 V/cm and injection time was controlled at 300 V/cm for 5 s. During all these procedures, electronic polar of CE would be reversed and LPA freshly replaced after using for every four times. P100-a 20-mer antisense PS-ODNs, contained primarily (n-1), (n-2), (n+1), and (n+2) deletion or appending sequences, had been separated with single-base resolution when using this polymer system. The separations of target PS-ODNs of P100 and P101, which was designed for anti-lung cancer, were investigated and their separation conditions were discussed here. The relative standard deviations of repeatability (n=6) between migration time and peak area were 0.4% and 0.5%, 0.5% and 0.5% for P100 and P101, respectively. Almost the same results would be observed for the relative standard deviations of intra-day and inter-day precision as well. These results showed that the capillary electrophoresis of LPA was very suitable for the separation and analysis of the PS-ODNs.