Fabrice Audouin

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

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 tert-butyl 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.

Functional Brush-Decorated Poly(globalide) Films by ARGET-ATRP for Bioconjugation

Formation of hydroxyl functionalized poly(macrolactone) films was achieved by a combination of enzymatic ring opening polymerization of globalide and thiol-ene coupling reactions. Reaction with α-bromoisobutyryl bromide yielded ATRP initiator functional films. Efficient activator regeneration by electron transfer (ARGET) ATRP was employed for the polymerization of tert-butyl acrylate (tBA) and NIPAM from the initiator decorated surfaces. Deprotection of the tert-butyl ester decorated polymer films yielded carboxylic acid groups for convenient attachment of biomolecule. The successful conjugation of green fluorescent protein (eGFP) and active enzyme chitobiase (Chb) on densely poly(acrylic acid)-functionalized polyester films is demonstrated.