Protein Immobilization onto Poly(acrylic acid) Functional Macroporous PolyHIPE Obtained by Surface-Initiated ARGET ATRP

Abstract

Amino-functional macroporous monoliths from polymerized high internal phase emulsion (polyHIPE) were surface modified with initiators for atom transfer radical polymerization (ATRP). The ATRP initiator groups on the polyHIPE surface were successfully used to initiate activator regeneration by electron transfer (ARGET) ATRP of (meth)acrylic monomers, such as methyl methacrylate (MMA) or tert-butyl acrylate (tBA) resulting in a dense coating of polymers on the polyHIPE surface. Addition of sacrificial initiator permitted control of the amount of polymer grafted onto the monolith surface. Subsequent removal of the tertbutyl protecting groups yielded highly functional polyHIPE-g-poly(acrylic acid). The versatility to use the high density of carboxylic acid groups for secondary reactions was demonstrated by the successful conjugation of enhanced green fluorescent protein (eGFP) and coral derived red fluorescent protein (DsRed) using EDC/sulfo-NHS chemistry, on the polymer 3D-scaffold surface. The materials and methodologies presented here are simple and robust, thus, opening new possibilities for the bioconjugation of highly porous polyHIPE for bioseparation applications.

Stable Poly(methacrylic acid) Brush Decorated Silica Nano-Particles by ARGET ATRP for Bioconjugation

“…Immobilization of biomolecules, such as proteins, enzymes, or antibodies on solid substrates has great importance in biosensors and biotechnology applications owing to their specific affinity for their targets [52,53]. Due to their robustness and versatility, functional polymer brushes have been utilized extensively for the immobilization of biomolecules [12,20,[52][53][54][55][56]. Herein, horseradish peroxidase (HRP) has been chosen to conceptually investigate the covalent immobilization through carboxylic acid groups of PMAA decorated Si-NP.…”

“…Very useful in that respect are polymer brushes, i.e., functional polymers attached to the nanoparticle surface, due to their high polymer segment density [10,11]. We are interested in the decoration of Si-NPs with poly(methacrylic acid) (PMAA) or poly(acrylic acid) (PAA) as they can easily be conjugated with biomolecules by conventional coupling chemistry [12]. Moreover, the hydrophilic nature of these brushes stabilizes the nano-particle suspension in aqueous media thereby preventing particle aggregation and ensuring extended shelf life [13].…”

“…In ARGET ATRP the amount of copper catalyst is significantly reduced by a strategy that employs a reducing agent to in situ regenerate the active Cu(I) species. This technique has successfully been employed from surfaces [12,[24][25][26][27][28] but to the best of our knowledge there are only very few reports of ARGET ATRP from Si-NP [29,30].…”

Optical formation of polymeric materials with heterogeneously distributed nanopores from a photopolymerizable composite

“…Such materials may be extensively used to produce various separation membranes [5][6][7][8], as microreactors to form nanoparticles of inorganic compounds [9], as a medium for liquid or gas chromatography [10], and as alternative dielectric layers with extra low dielectric permeability in high-performance microelectronics [11]. In addition, the pore surface can be modified by functional groups or active sites to impart the desired properties of the porous materials [2,[6][7][8].…”

“…Such materials may be extensively used to produce various separation membranes [5][6][7][8], as microreactors to form nanoparticles of inorganic compounds [9], as a medium for liquid or gas chromatography [10], and as alternative dielectric layers with extra low dielectric permeability in high-performance microelectronics [11]. In addition, the pore surface can be modified by functional groups or active sites to impart the desired properties of the porous materials [2,[6][7][8]. This approach allows a number of problems to be solved that are associated with heterogeneous catalysis control [12], new media creation for chromatography [2,6], and the making of nanofilters with selective separation of complicated water-organic mixtures [7,8].…”