Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.
Identifieur interne : 002812 ( PubMed/Corpus ); précédent : 002811; suivant : 002813Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.
Auteurs : G J Bruin ; K O Börnsen ; D. Hüsken ; E. Gassmann ; H M Widmer ; A. PaulusSource :
- Journal of chromatography. A [ 0021-9673 ] ; 1995.
English descriptors
- KwdEn :
- 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
Links to Exploration step
pubmed:7581844Le document en format XML
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Gassmann</name></author><author><name sortKey="Widmer, H M" sort="Widmer, H M" uniqKey="Widmer H" first="H M" last="Widmer">H M Widmer</name></author><author><name sortKey="Paulus, A" sort="Paulus, A" uniqKey="Paulus A" first="A" last="Paulus">A. Paulus</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1995">1995</date><idno type="RBID">pubmed:7581844</idno><idno type="pmid">7581844</idno><idno type="doi">10.1016/0021-9673(95)00231-b</idno><idno type="wicri:Area/PubMed/Corpus">002812</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002812</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.</title><author><name sortKey="Bruin, G J" sort="Bruin, G J" uniqKey="Bruin G" first="G J" last="Bruin">G J Bruin</name><affiliation><nlm:affiliation>Corporate Analytical Research, Ciba, Basel, Switzerland.</nlm:affiliation></affiliation></author><author><name sortKey="Bornsen, K O" sort="Bornsen, K O" uniqKey="Bornsen K" first="K O" last="Börnsen">K O Börnsen</name></author><author><name sortKey="Husken, D" sort="Husken, D" uniqKey="Husken D" first="D" last="Hüsken">D. Hüsken</name></author><author><name sortKey="Gassmann, E" sort="Gassmann, E" uniqKey="Gassmann E" first="E" last="Gassmann">E. Gassmann</name></author><author><name sortKey="Widmer, H M" sort="Widmer, H M" uniqKey="Widmer H" first="H M" last="Widmer">H M Widmer</name></author><author><name sortKey="Paulus, A" sort="Paulus, A" uniqKey="Paulus A" first="A" last="Paulus">A. Paulus</name></author></analytic><series><title level="j">Journal of chromatography. A</title><idno type="ISSN">0021-9673</idno><imprint><date when="1995" type="published">1995</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>Electrophoresis (methods)</term><term>Endonucleases (chemistry)</term><term>Mass Spectrometry (methods)</term><term>Molecular Sequence Data</term><term>Oligonucleotides, Antisense (chemistry)</term><term>Poly T (chemistry)</term><term>Snake Venoms (enzymology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Endonucleases</term><term>Oligonucleotides, Antisense</term><term>Poly T</term></keywords><keywords scheme="MESH" type="chemical" qualifier="enzymology" xml:lang="en"><term>Snake Venoms</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Electrophoresis</term><term>Mass Spectrometry</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Molecular Sequence Data</term></keywords></textClass></profileDesc></teiHeader><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></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">7581844</PMID><DateCompleted><Year>1995</Year><Month>12</Month><Day>05</Day></DateCompleted><DateRevised><Year>2019</Year><Month>07</Month><Day>20</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0021-9673</ISSN><JournalIssue CitedMedium="Print"><Volume>709</Volume><Issue>1</Issue><PubDate><Year>1995</Year><Month>Aug</Month><Day>11</Day></PubDate></JournalIssue><Title>Journal of chromatography. A</Title><ISOAbbreviation>J Chromatogr A</ISOAbbreviation></Journal><ArticleTitle>Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.</ArticleTitle><Pagination><MedlinePgn>181-95</MedlinePgn></Pagination><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Bruin</LastName><ForeName>G J</ForeName><Initials>GJ</Initials><AffiliationInfo><Affiliation>Corporate Analytical Research, Ciba, Basel, Switzerland.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Börnsen</LastName><ForeName>K O</ForeName><Initials>KO</Initials></Author><Author ValidYN="Y"><LastName>Hüsken</LastName><ForeName>D</ForeName><Initials>D</Initials></Author><Author ValidYN="Y"><LastName>Gassmann</LastName><ForeName>E</ForeName><Initials>E</Initials></Author><Author ValidYN="Y"><LastName>Widmer</LastName><ForeName>H M</ForeName><Initials>HM</Initials></Author><Author ValidYN="Y"><LastName>Paulus</LastName><ForeName>A</ForeName><Initials>A</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>Netherlands</Country><MedlineTA>J Chromatogr A</MedlineTA><NlmUniqueID>9318488</NlmUniqueID><ISSNLinking>0021-9673</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D016376">Oligonucleotides, Antisense</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012910">Snake Venoms</NameOfSubstance></Chemical><Chemical><RegistryNumber>25086-81-1</RegistryNumber><NameOfSubstance UI="D011071">Poly T</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 3.1.-</RegistryNumber><NameOfSubstance UI="D004720">Endonucleases</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004586" MajorTopicYN="N">Electrophoresis</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="Y">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004720" MajorTopicYN="N">Endonucleases</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013058" MajorTopicYN="N">Mass Spectrometry</DescriptorName><QualifierName UI="Q000379" MajorTopicYN="N">methods</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008969" MajorTopicYN="N">Molecular Sequence Data</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D016376" MajorTopicYN="N">Oligonucleotides, Antisense</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011071" MajorTopicYN="N">Poly T</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012910" MajorTopicYN="N">Snake Venoms</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="N">enzymology</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>1995</Year><Month>8</Month><Day>11</Day></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>1995</Year><Month>8</Month><Day>11</Day><Hour>0</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>1995</Year><Month>8</Month><Day>11</Day><Hour>0</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">7581844</ArticleId><ArticleId IdType="pii">0021-9673(95)00231-B</ArticleId><ArticleId IdType="doi">10.1016/0021-9673(95)00231-b</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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