MersV1, Main, Exploration, bibRecord, 004023

Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

Identifieur interne : 004023 ( Main/Exploration ); précédent : 004022; suivant : 004024

Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

Auteurs : G J Bruin [Suisse] ; K O Börnsen ; D. Hüsken ; E. Gassmann ; H M Widmer ; A. Paulus

Source :

Journal of chromatography. A [ 0021-9673 ] ; 1995.

RBID : pubmed:7581844

Descripteurs français

KwdFr :
- Données de séquences moléculaires, Endonucleases (), Oligonucléotides antisens (), Poly T (), Spectrométrie de masse (), Séquence nucléotidique, Venins de serpent (enzymologie), Électrophorèse ().
MESH :
- enzymologie : Venins de serpent.
- Données de séquences moléculaires, Endonucleases, Oligonucléotides antisens, Poly T, Spectrométrie de masse, Séquence nucléotidique, Électrophorèse.

English descriptors

KwdEn :
- Base Sequence, Electrophoresis (methods), Endonucleases (chemistry), Mass Spectrometry (methods), Molecular Sequence Data, Oligonucleotides, Antisense (chemistry), Poly T (chemistry), Snake Venoms (enzymology).
MESH :
- chemical , chemistry : Endonucleases, Oligonucleotides, Antisense, Poly T.
- chemical , enzymology : Snake Venoms.
- methods : Electrophoresis, Mass Spectrometry.
- Base Sequence, Molecular Sequence Data.

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.

DOI: 10.1016/0021-9673(95)00231-b
PubMed: 7581844

Affiliations:

Suisse

Le document en format XML

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<author><name sortKey="Gassmann, E" sort="Gassmann, E" uniqKey="Gassmann E" first="E" last="Gassmann">E. Gassmann</name>
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<author><name sortKey="Widmer, H M" sort="Widmer, H M" uniqKey="Widmer H" first="H M" last="Widmer">H M Widmer</name>
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<term>Molecular Sequence Data</term>
<term>Oligonucleotides, Antisense (chemistry)</term>
<term>Poly T (chemistry)</term>
<term>Snake Venoms (enzymology)</term>
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<term>Endonucleases ()</term>
<term>Oligonucléotides antisens ()</term>
<term>Poly T ()</term>
<term>Spectrométrie de masse ()</term>
<term>Séquence nucléotidique</term>
<term>Venins de serpent (enzymologie)</term>
<term>Électrophorèse ()</term>
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<keywords scheme="MESH" type="chemical" qualifier="enzymology" xml:lang="en"><term>Snake Venoms</term>
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<term>Mass Spectrometry</term>
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<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>Molecular Sequence Data</term>
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<term>Oligonucléotides antisens</term>
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<term>Spectrométrie de masse</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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<country name="Suisse"><noRegion><name sortKey="Bruin, G J" sort="Bruin, G J" uniqKey="Bruin G" first="G J" last="Bruin">G J Bruin</name>
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Serveur d'exploration MERS

Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

	Serveur d'exploration MERS
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