STABILITY MEASUREMENTS OF ANTISENSE OLIGONUCLEOTIDES BY CAPILLARY GEL-ELECTROPHORESIS

被引：29

作者：

BRUIN, GJM ^{[1
]}

BORNSEN, KO ^{[1
]}

HUSKEN, D ^{[1
]}

GASSMANN, E ^{[1
]}

WIDMER, HM ^{[1
]}

PAULUS, A ^{[1
]}

机构：

[1] CIBA GEIGY AG, CENT RES LABS, CH-4002 BASEL, SWITZERLAND

来源：

JOURNAL OF CHROMATOGRAPHY A | 1995年 / 709卷 / 01期

关键词：

D O I：

10.1016/0021-9673(95)00231-B

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The approach of using antisense oligonucleotides as potential drugs is based on hybridization of a short chemically-modified oligonucleotide with complementary cellular DNA or RNA sequences. A critical question is the stability of chemically modified antisense oligonucleotides in cellular environments. In a model system, resistance against various nucleases was evaluated by capillary gel electrophoresis (CGE). For some of the samples, matrix assisted laser desorption and ionization mass spectrometry (MALDI-MS) was used as an additional analytical tool to perform stability measurements. Using CGE, the enzymatic degradation of single nucleotides from the oligomer can be followed after different incubation times. 10% T polyacrylamide gels give baseline resolution for oligonucleotides ranging between 5 and 30 bases in length. The kinetic influence of a specific nuclease concentration and the antisense oligonucleotide structure on the cleavage reaction are discussed. Also, a simple desalting method to improve the injection efficiency and sensitivity of the method are described. Examples of measurements of chemically modified antisense 19-mers are presented.

引用

页码：181 / 195

页数：15

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