Reactive polymers in diagnostics: Syntheses and characterizations of nucleic acid probes and maleic anhydride-co-methyl vinyl ether polymers
✍ Scribed by C. Ladavière; L. Veron; T. Delair; A. Domard; C. Pichot; B. Mandrand
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 256 KB
- Volume
- 65
- Category
- Article
- ISSN
- 0021-8995
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✦ Synopsis
Maleic anhydride-co-methyl vinyl ether copolymers were used as reactive polymers to link oligodeoxyribonucleotides (ODN) to make oligonucleotide-copolymer conjugates of potential applications in diagnostics. The anhydride moieties of the copolymers were used for the covalent binding, via the formation of a peptide bond, on reaction with DNA probes that were amino modified at the 5 position. The best coupling yields were obtained by carrying out the reaction in a DMSO-borate buffer (95/5 volume ratio) mixture at high ionic strength. Copolymers and ODN-copolymer conjugates were characterized by size exclusion chromatography and multiangle laser light scattering detection. The average molecular weights of the formed conjugates were relatively high, in the 10 6 g/mol range, resulting from the crosslinking of several copolymer molecules during coupling. Coupling DNA probes onto smaller molecular weight polymers did not result in the formation of very high molecular weight conjugates showing that copolymer chain interpenetration was a determining factor of the crosslinking process that occurs during the coupling reaction.