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Synthesis and characterization of nucleic acid–(co)polymer conjugates: application to diagnostics

Identifieur interne : 001330 ( Istex/Checkpoint ); précédent : 001329; suivant : 001331

Synthesis and characterization of nucleic acid–(co)polymer conjugates: application to diagnostics

Auteurs : Thierry Delair [France] ; Marie-Hélène Charles [France] ; Philippe Cros [France] ; Ali Laayoun [France] ; Bernard Mandrand [France] ; Christian Pichot [France]

Source :

RBID : ISTEX:2978E7AC880E2D756C45A9EEB5082091302760D2

English descriptors

Abstract

This paper provides a survey of the synthesis of oligodeoxyribonucleotides (ODNs)–(co)polymer conjugates for application to diagnostics. Reactive copolymers containing various chemical functions (carboxylic acid groups under the form of activated esters or aldehyde) were used to covalently link ODNs. It was found that this immobilization reaction was dependent upon several parameters including the chemical reactivity of the functional group, the structural parameters (conformation) of the polymer, and the hydrogen bonding capability between the two macromolecular partners. Analysis of the conjugation products by size exclusive chromatography – light scattering displayed higher molecular weights than expected because of a concomitant aggregation process resulting from either hydrogen bonding between ODNs and polymer chains or side reactions from heterocyclic base amino groups. The extent of this aggregation process was dependent on the chemical nature of the polymer and base composition of the probes and it could be partly or totally avoided upon decreasing the capacity of the nucleic acids to form hydrogen bonds. These conjugates were tested in nucleic acid detection and they were shown to improve assay in both capture and signal efficiency for a long double‐stranded DNA. © 1998 John Wiley & Sons, Ltd.

Url:
DOI: 10.1002/(SICI)1099-1581(199806)9:6<349::AID-PAT789>3.0.CO;2-J


Affiliations:


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ISTEX:2978E7AC880E2D756C45A9EEB5082091302760D2

Le document en format XML

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<name sortKey="Charles, Marie Elene" sort="Charles, Marie Elene" uniqKey="Charles M" first="Marie-Hélène" last="Charles">Marie-Hélène Charles</name>
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<term>Aggregate formation</term>
<term>Aggregated</term>
<term>Aggregated conjugates</term>
<term>Aggregation</term>
<term>Aggregation phenomenon</term>
<term>Aggregation process</term>
<term>Aldehyde</term>
<term>Aldehyde groups</term>
<term>Aldehydic polymer</term>
<term>Aldehydic polymers</term>
<term>Amine</term>
<term>Amino groups</term>
<term>Amplification</term>
<term>Background signal</term>
<term>Base composition</term>
<term>Base composition effect</term>
<term>Base effect</term>
<term>Basic principle</term>
<term>Bioconjugate chem</term>
<term>Borate buffer</term>
<term>Broad peak</term>
<term>Carboxylic acid groups</term>
<term>Chemical nature</term>
<term>Chemical reactivity</term>
<term>Conjugate</term>
<term>Conjugation</term>
<term>Copolymer</term>
<term>Covalent</term>
<term>Covalent bonds</term>
<term>Covalent immobilization</term>
<term>Delair</term>
<term>Detection conjugate</term>
<term>Detection conjugates</term>
<term>Detection limit</term>
<term>Elosa test</term>
<term>Elosa tests</term>
<term>Ester</term>
<term>Exclusion limit</term>
<term>Experimental conditions</term>
<term>Form hydrogen bonds</term>
<term>Free solution capillary electrophoresis</term>
<term>Functional groups</term>
<term>Heterocyclic base</term>
<term>High molar mass</term>
<term>Homogeneous sequences</term>
<term>Hybridization</term>
<term>Hydrogen bonds</term>
<term>Hydrolysis reaction</term>
<term>Hydrolysis reaction rate</term>
<term>Immobilization</term>
<term>Immobilization reaction</term>
<term>Immobilization yields</term>
<term>Ionic strength</term>
<term>John wiley sons</term>
<term>Laser light</term>
<term>Less aggregated material</term>
<term>Maleic anhydride</term>
<term>Mandrand</term>
<term>Methyl vinyl ether</term>
<term>Molecular weight</term>
<term>Molecular weights</term>
<term>More details</term>
<term>Nabh4 reduction</term>
<term>Noise ratio</term>
<term>Nonspecific adsorption</term>
<term>Nucleic</term>
<term>Nucleic acid conjugates</term>
<term>Nucleic acid detection</term>
<term>Nucleic acid probes</term>
<term>Nucleic acids</term>
<term>Nucleophilic attack</term>
<term>Odns</term>
<term>Oligodeoxyribonucleotides</term>
<term>Oligonucleotide</term>
<term>Oligonucleotide conjugates</term>
<term>Oligonucleotide probes</term>
<term>Oligonucleotides</term>
<term>Peak shapeb</term>
<term>Phosphoramidite chemistry</term>
<term>Physicochemical properties</term>
<term>Pichot</term>
<term>Polym</term>
<term>Polymer</term>
<term>Polymer chain</term>
<term>Polymer chains</term>
<term>Primary amine</term>
<term>Probe</term>
<term>Reactive</term>
<term>Reactive copolymers</term>
<term>Reactive groups</term>
<term>Reactive polymers</term>
<term>Reactivity ratios</term>
<term>Reductive amination</term>
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<term>Saccharidic copolymer</term>
<term>Saccharidic polymer</term>
<term>Saccharidic polymers</term>
<term>Same time</term>
<term>Sharp decrease</term>
<term>Side reactions</term>
<term>Signal amplification</term>
<term>Size exclusion chromatography</term>
<term>Solid phase</term>
<term>Solid support</term>
<term>Specific signals</term>
<term>Synthetic carrier</term>
<term>Unpublished results</term>
<term>Various approaches</term>
<term>Various biomolecules</term>
<term>Various conjugates</term>
<term>Various factors</term>
<term>Various reactive polymers</term>
<term>Water molecules</term>
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<div type="abstract" xml:lang="en">This paper provides a survey of the synthesis of oligodeoxyribonucleotides (ODNs)–(co)polymer conjugates for application to diagnostics. Reactive copolymers containing various chemical functions (carboxylic acid groups under the form of activated esters or aldehyde) were used to covalently link ODNs. It was found that this immobilization reaction was dependent upon several parameters including the chemical reactivity of the functional group, the structural parameters (conformation) of the polymer, and the hydrogen bonding capability between the two macromolecular partners. Analysis of the conjugation products by size exclusive chromatography – light scattering displayed higher molecular weights than expected because of a concomitant aggregation process resulting from either hydrogen bonding between ODNs and polymer chains or side reactions from heterocyclic base amino groups. The extent of this aggregation process was dependent on the chemical nature of the polymer and base composition of the probes and it could be partly or totally avoided upon decreasing the capacity of the nucleic acids to form hydrogen bonds. These conjugates were tested in nucleic acid detection and they were shown to improve assay in both capture and signal efficiency for a long double‐stranded DNA. © 1998 John Wiley & Sons, Ltd.</div>
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