Khan K, Van Schepdael A, Saison-Behmoaras T, Van Aerschot A, Hoogmartens J
Laboratorium voor Farmaceutische Chemie en Analyse van Geneesmiddelen, Faculteit Farmaceutische Wetenschappen, K.U. Leuven, Belgium. kamran=khan%fca%
Electrophoresis. 1998 Sep;19(12):2163-8. doi: 10.1002/elps.1150191220.
An attempt was made to evaluate the stability of an antisense oligonucleotide against nucleases present in HBL 100ras cells. To detect nanomolar concentrations of the oligonucleotide, a sensitive detection system was required. A combination of capillary electrophoresis/laser-induced fluorescence (CE-LIF) with fluorescence derivatization did not improve the sensitivity significantly and also resulted in loss of separation of the derivatized sample. On-column isotachophoresis for the preconcentration of oligonucleotide samples in DB-17 coated capillaries filled with hydroxyethyl cellulose solution could be an alternative. The isotachophoresis (ITP) step allows injection of up to 40% of the capillary volume without loss in peak resolution and peak efficiency. Using ITP-capillary polymer sieving electrophoresis (CPSE), the limit of quantitation at a signal-to-noise ratio of 10 was 73 ng/mL for a 12-mer oligonucleotide. Using these conditions, the gain in sensitivity was 125.
研究人员尝试评估一种反义寡核苷酸对HBL 100ras细胞中存在的核酸酶的稳定性。为了检测纳摩尔浓度的寡核苷酸，需要一个灵敏的检测系统。毛细管电泳/激光诱导荧光（CE-LIF）与荧光衍生化相结合的方法并没有显著提高灵敏度，而且还导致衍生化样品的分离度下降。在填充有羟乙基纤维素溶液的DB-17涂层毛细管中，通过柱上等速电泳对寡核苷酸样品进行预浓缩可能是一种替代方法。等速电泳（ITP）步骤允许注入高达40%的毛细管体积，而不会损失峰分辨率和峰效率。使用ITP-毛细管聚合物筛分电泳（CPSE），对于12聚体寡核苷酸，在信噪比为10时的定量限为73 ng/mL。在这些条件下，灵敏度提高了125倍。
