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

Abstract

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.

Making the best of it: nitroxide-mediated polymerization of methacrylatesviathe copolymerization approach with functional styrenics

“…These structures span a relatively large scope of functionality and reactivity, as shown in Scheme 2. Only Sty had previously been employed with the sole purpose of gaining control over the NMP of methacrylic esters, 31,33,[53][54][55][56][57][58][59][60][61] while PFS had been shown to exert a positive influence as well, yet in a study not focusing on the control aspect.…”

“…POEGMA is particularly interesting as it is frequently employed in the biomaterials field, due to demonstrated biocompatibility and ease of synthesis. 37,69 It was already polymerized by NMP following the copolymerization approach, using Sty, 54,57 acrylonitrile, 37,70 PFS, 43 and MPDL. 44 The same type of kinetic experiments as for MMA was therefore undertaken.…”

Reactive and Functional Nanoobjects by Polymerization‐Induced Self‐Assembly

“…Schematic representation of the synthesis of NTA‐functionalized polymeric nanoparticles by NMP and NMPISA, followed by nickel‐mediated immobilization of His‐tagged enzymes on their surface. Reproduced with permission . Copyright 2017, American Chemical Society.…”

“…For instance, Keller et al synthesized nitrilotriacetic acid (NTA)-displaying nanoparticles by nitroxide-mediated PISA (NMPISA; Scheme 12). [67] A protected NTA alkoxyamine initiator (pNTA-MAMA-SG1; see Scheme 11) was first designed and employed for the homopoly merization of long OEGMA (M n 950 g mol −1 ). This NTA-functionalized macroinitiator was then involved in the NMPISA of n-butyl methacrylate (BMA).…”

ICAR ATRP in PEG with Low Concentration of Cu(II) Catalyst: A Versatile Method for Synthesis of Block Copolymer Nanoassemblies under Dispersion Polymerization

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radical polymerization including reversible addition fragmentation transfer (RAFT) polymerization, [4][5][6][7][8][9][10][11][12][13][14][15][16] atom transfer radical polymerization (ATRP) [17][18][19][20][21] and nitroxide-mediated polymerization [22,23] have been adopted in PISA. Of all these, PISA adopting RAFT under dispersion polymerization, for example, macro-RAFT agent-mediated dispersion polymerization, is the most widely reported one, [5][6][7][8][9][10][11][12][13][14][15][16] and various block copolymer nanoassemblies have been synthesized by aqueous RAFT dispersion polymerization or nonaqueous RAFT dispersion polymerization.

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