Oligonucleotide synthesis on maleic anhydride copolymers covalently bound to silica spherical support and characterization of the obtained conjugates

Abstract

ABSTRACT:A new route was proposed to make polymer-oligonucleotide conjugates of potential applications in diagnostics. It consisted in direct synthesis of oligonucleotides onto controlled pore glass surface grafted with poly(maleic anhydride-alt-methyl vinyl ether) (P[MAMVE]) or poly(maleic anhydride-alt-ethylene) (P [MAE]). The anhydride moieties were used for both the covalent coupling of the copolymer via ester bond and binding of 5Ј-dimethoxytrityl thymidine 3Ј-(6-aminohexyl phosphate) (I) initiator of oligodeoxynucleotide (ODN) synthesis via amide bond. The difference of stability between ester and amide links under basic treatment was used for the selective cleavage of (polymer-oligonucleotide) conjugates after DNA synthesis completion. We succeeded in grafting functionalized copolymer onto silica surface and Polythymidine 26-mer ODN was performed. After concentrated ammonium hydroxide treatment, conjugate crude materials were characterized by size exclusion chromatography coupled to multiangle laser light scattering detection. The number average molecular weight (Mn) for conjugate with P[MAMVE] was abnormally lower than expected and was assigned to polymer degradation using high pH conditions. Such a phenomenon did not occur with P[MAE]-polythymidine conjugate. However, in both cases, parasite ODN synthesis was also evidenced, which was attributed to thymidine phosphoramidite adsorption side reaction during DNA synthesis.

Bioengineering functional copolymers. XII. Interaction of boron-containing and PEO branched derivatives of poly(MA-alt-MVE) with HeLa cells

“…It is known that these copolymers can be regarded as pre-activated polymers due to the presence of anhydride moieties susceptible to the reaction with a primary amine of a biomolecule [9]. The alternating copolymers of maleic anhydride (MA) with methyl vinyl ether (MVE) or divinyl ether (DVE) were utilized in various applications in diagnostics [10,11] and in chemotherapy as effective antitumor agents [8]. Poly (MA-alt-DVE), known as pyran copolymer is one of the well known bioengineering polymers having a wide range of biological activity.…”

Bioengineering Functional Copolymers. IX. Poly[(maleic anhydride‐co‐hexene‐1)‐g‐poly(ethylene oxide)]

“…[10,11] These copolymers and their functionalized derivatives are used as precursors for enzyme synthesis, [12] as biologically active agents against several transplantable tumors (the half-amide-half-carboxylate salt of the alternating copolymer of MA with ethylene), [13] and for the preparation of exchange membrane systems, adhesives, and coatings. [1,2] The oligonucleotide synthesis of MA copolymers covalently bound to a silica spherical support and the characterization of the obtained conjugates have been described by Chaix et al [14] Direct synthesis of oligonucleotides was achieved on a controlled porous glass surface grafted with alternating copolymers of MA, including poly(MA-alt-ethylene).…”

Nucleic Acid/Organic Polymer Hybrid Materials: Synthesis, Superstructures, and Applications

“…Another possibility to generate DNA side-chain polymers on the solid phase relied on immobilizing the synthetic polymer backbone on glass beads. [108] The hydroxy groups present on the controlled pore glass surface were reacted with alternating copolymers containing maleic anhydride units so that a covalent ester bond between the support and the polymer formed. The remaining anhydride structures were utilized to bind a 5'-trityl-protected mononucleotide carrying an aminoalkyl spacer by means of bond formation that acted as an initiator of ODN synthesis.…”