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Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

Bruin, Gerardus and Börnsen, K O and Hüsken, D and Gassmann, Ernst and Widmer, H M and Paulus, A (1995) Stability measurements of antisense oligonucleotides by capillary gel electrophoresis. Journal of chromatography. A, 709 (1). pp. 181-195. ISSN 0021-9673

Abstract

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.

Item Type: Article
Date Deposited: 31 Jan 2012 00:45
Last Modified: 01 Feb 2013 00:45
URI: https://oak.novartis.com/id/eprint/6538

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