Bioconjugation of D‐glucuronic acid sodium salt to well‐defined primary amine‐containing homopolymers and block copolymers

Abstract

The synthesis of well-defined carboxylic acid-functionalized glycopolymers prepared via one-step postpolymerization modification of poly(N-[3-aminopropyl] methacrylamide) (PAPMA), a water-soluble primary amine methacrylamide, in aqueous medium is demonstrated. PAPMA was first polymerized via aqueous reversible addition-fragmentation chain transfer polymerization in aqueous buffer using 4-cyanopentanoic acid dithiobenzoate as the chain transfer agent and 4,4 0 -azobis(4-cyanovaleric acid) (V-501) as the initiator at 70 C. The resulting well-defined PAPMA was then conjugated with D-glucuronic acid sodium salt through reductive amination in alkaline medium (pH 8.5) at 45 C. The successful bioconjugation was proven through proton ( 1 H) and carbon ( 13 C) nuclear magnetic resonance spectroscopy and matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis, which indicated near quantitative conversion. A similar bioconjugation reaction was conducted with poly(2-aminoethyl methacrylate) (PAEMA) and poly(2-aminoethyl methacrylate-b-poly(N-[2hydroxypropyl]methacrylamide) (PAEMA-b-PHPMA). For the PAEMA homopolymers and block copolymers, however, lower conversion was obtained, most likely because of degradation reactions of PAEMA in alkaline medium. V C 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 48: 3052-3061, 2010

Use of the interior cavity of the P22 capsid for site-specific initiation of atom-transfer radical polymerization with high-density cargo loading

“…According to this method the radius of the particles remains nearly constant, measuring at 29.3 nm (P22 S39C -int) and 28.2 nm (P22 S39C -xAEMA), while the molecular weight increases upon polymerization. Using the standard dn/dc for protein (0.185) and the published value for AEMA (0.153) 27 the multi-angle light scattering data from the P22 S39C -int and P22 S39C -xAEMA samples were fit to obtain molecular weights. The measured molecular weight for P22 S39C -int was 18.9 ± 0.2 MDa, consistent with the predicted value for the EX morphology of P22, while the P22 S39C -xAEMA had a combined molecular weight of 20.2 ± 0.6 MDa, with 18.7 ± 0.5 MDa contributed from protein and 1.5 ± 0.4 MDa contributed from polymer.…”

“…Using the elution profile data the number average molecular weight, Mn, was calculated with Astra 5.3.14 software (Wyatt Technologies) using a dn/dc for protein of 0.185 and a dn/dc for the polymer component of 0.153. 27 …”

Synthesis of Glycopolymer Architectures by Reversible-Deactivation Radical Polymerization

“…Post-polymerization conjugation of D-glucuronic acid to water soluble polymer featuring primary amino functions as reported by Alidedeoglu et al [211].…”

“…Alidedeoglu et al [211] synthesized well-defined glycopolymers carrying 1-amino-1-deoxy-alditol residues derived from D-glucuronic acid directly in water (Entry 256-258, Table 3). To this end, 2-aminoethyl methacrylate M54 and 2-aminopropyl methacrylamide M55 were homopolymerized in acetic buffer (pH 5) at 50 °C and 70 °C, respectively, for around an hour in the presence of RAFT agent R1; the double hydrophilic copolymer poly(M54)-block-poly(M56) was also prepared under similar conditions.…”

RAFT polymerization mediated bioconjugation strategies

“…In studies by York et al 84,85 and Alidedeoglu et al 100 homoand copolymers of a primary amine containing monomer, N-(3-aminopropyl)methacrylamide (APMA) were prepared in an aqueous acetic buffer (pH 5.2 to prevent degradation of dithiobenzoate functionality) using 4-cyanopentanoic acid dithiobenzoate as a RAFT agent. APMA provided amine functionality, allowing conjugation of cell-targeting folate derivatives and o-glucuronic acid.…”