title: Protein microarrays based on polymer brushes prepared via surface-initiated atom transfer radical polymerization.
abstract: Polymer brushes represent an interesting platform for the development of high-capacity protein binding surfaces. Whereas the protein binding properties of polymer brushes have been investigated before, this manuscript evaluates the feasibility of poly(glycidyl methacrylate) (PGMA) and PGMA-co-poly(2-(diethylamino)ethyl methacrylate) (PGMA-co-PDEAEMA) (co)polymer brushes grown via surface-initiated atom transfer radical polymerization (SI-ATRP) as protein reactive substrates in a commercially available microarray system using tantalum-pentoxide-coated optical waveguide-based chips. The performance of the polymer-brush-based protein microarray chips is assessed using commercially available dodecylphosphate (DDP)-modified chips as the benchmark. In contrast to the 2D planar, DDP-coated chips, the polymer-brush-covered chips represent a 3D sampling volume. This was reflected in the results of protein immobilization studies, which indicated that the polymer-brush-based coatings had a higher protein binding capacity as compared to the reference substrates. The protein binding capacity of the polymer-brush-based coatings was found to increase with increasing brush thickness and could also be enhanced by copolymerization of 2-(diethylamino)ethyl methacrylate (DEAEMA), which catalyzes epoxide ring-opening of the glycidyl methacrylate (GMA) units. The performance of the polymer-brush-based microarray chips was evaluated in two proof-of-concept microarray experiments, which involved the detection of biotin-streptavidin binding as well as a model TNFα reverse assay. These experiments revealed that the use of polymer-brush-modified microarray chips resulted not only in the highest absolute fluorescence readouts, reflecting the 3D nature and enhanced sampling volume provided by the brush coating, but also in significantly enhanced signal-to-noise ratios. These characteristics make the proposed polymer brushes an attractive alternative to commercially available, 2D microarray surface coatings.
authors.affiliation: École Polytechnique Fédérale de Lausanne (EPFL), Institut des Matériaux and Institut des Sciences et Ingénierie Chimiques, Laboratoire des Polymères, Bâtiment MXD, Station 12, CH-1015 Lausanne, Switzerland.
authors.forename: Raphael
authors.initials: R
authors.lastname: Barbey
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Kauffmann
authors.forename: Markus
authors.initials: M
authors.lastname: Ehrat
authors.forename: Harm-Anton
authors.initials: HA
authors.lastname: Klok
mesh_terms: D001426:Bacterial Proteins; D001710:Biotin / Q000031:analogs & derivatives; D004852:Epoxy Compounds; D008689:Methacrylates; D009757:Nylons; D058105:Polymerization; D011108:Polymers / Q000737:chemistry*; D040081:Protein Array Analysis; D014409:Tumor Necrosis Factor-alpha
chemical_list: D001426:Bacterial Proteins; D004852:Epoxy Compounds; D008689:Methacrylates; D009757:Nylons; D011108:Polymers; D014409:Tumor Necrosis Factor-alpha; C066114:biotin-streptavidin complex; C109846:poly(2-(diethylamino)ethyl methacrylate); D001710:Biotin; C007870:glycidyl methacrylate