3 years ago

Nitrilotriacetic Amine-Functionalized Polymeric Core–Shell Nanoparticles as Enzyme Immobilization Supports

Nitrilotriacetic Amine-Functionalized Polymeric Core–Shell Nanoparticles as Enzyme Immobilization Supports
Manuel Ferrer, Dominic Keller, Mónica Martínez-Martínez, Ana Beloqui, Guillaume Delaittre
Nitrilotriacetic amine (NTA)-functionalized nanoparticles obtained by aqueous polymerization-induced self-assembly (PISA) are introduced as immobilization supports for polyhistidine-functionalized (His-tagged) enzymes. A novel initiator for nitroxide-mediated polymerization based on the nitroxide SG1 and carrying a protected NTA moiety was first synthesized. Size-exclusion chromatography (SEC) and electrospray ionization mass spectrometry (ESI-MS) proved the ability of this initiator to produce well-defined end-functional vinyl polymers. Subsequently, oligo(ethylene glycol) methacrylate-based macroinitiators were synthesized and chain-extended to form amphiphilic block copolymer nanoparticles, either by nanoprecipitation or by PISA. The latter method yielded spherical nanoparticles with a higher definition, as demonstrated by dynamic light scattering (DLS). Deprotection of the NTA moiety and complexation with nickel ions were assessed by DLS and inductively coupled plasma optical emission spectroscopy/mass spectrometry (ICP-OES/MS). Finally, immobilization of His-tagged horseradish peroxidase and ester hydrolase were successfully carried out, leading to catalytically active nanobiocatalysts, as shown by UV–vis measurements.

Publisher URL: http://dx.doi.org/10.1021/acs.biomac.7b00677

DOI: 10.1021/acs.biomac.7b00677

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