A novel metal-based chelating method has been used to provide an order of magnitude increase in immunoassay performance on COC plastics compared to passive binding. COCs are hydrophobic and without surface modification are often unsuitable for applications where protein adhesion is desired. When interacting with the bare plastic, the majority of the bound proteins will be denatured and become non-functional. Many of the surface modification techniques reported to-date require costly equipment setup or the use of harsh reaction conditions. Here, we have successfully demonstrated the use of a simple and quick metal chelation method to increase the sensitivity, activity and efficiency of protein binding to COC surfaces. A detailed analysis of the COC surfaces after activation with the metal complexes is presented, and the immunoassay performance was studied using three different antibody pairs.
[Show abstract][Hide abstract] ABSTRACT: Using a high-throughput surface discovery approach, we have generated a 1600-member library of metal-containing surfaces and screened them for antibody binding potential. The surface library assembly involved graft modification of argon plasma-treated polyvinylidenedifluoride (PVDF) membranes with alternating maleic anhydride-styrene copolymer followed by anhydride ring opening with a range of secondary amines and microarray contact printing of transition metal complexes. The microarrays of metal-containing surfaces were then tested for their antibody binding capacity by incubation with a biotinylated mouse antibody in a chemiluminescence assay. A total of 11 leads were identified from the first screen, constituting a "hit" rate of 0.7%. A smaller 135-member surface library was then synthesized and screened to optimize existing hits and generate additional leads. To demonstrate the applicability of these surfaces to other formats, high-binding surface leads were then transferred onto Luminex beads for use in a bead flow cytometric immunoassay. The novel one-step antibody coupling process increased assay sensitivity of a Luminex tumor necrosis factor immunoassay. These high-binding surfaces do not require prior incorporation of polyhistidine tags or posttreatments such as oxidation to achieve essentially irreversible binding of immunoglobulin G.
[Show abstract][Hide abstract] ABSTRACT: We present what is believed to be the first microstructured polymer optical fiber (mPOF) fabricated from Topas cyclic olefin copolymer, which has attractive material and biochemical properties. This polymer allows for a novel type of fiber-optic biosensor, where localized sensor layers may be activated on the inner side of the air holes in a predetermined section of the mPOF. The concept is demonstrated using a fluorescence-based method for selective detection of fluorophore-labeled antibodies. (c) 2007 Optical Society of America.
[Show abstract][Hide abstract] ABSTRACT: Nanoparticles and their interaction with human cells have been a focus of many groups during the past decade. We discuss and review here the progress in the field of understanding and harnessing the interactions of polymeric nanoparticles synthesized by the miniemulsion process with different cell types. Nanotechnology and the hereby produced nanomaterials have promised to make use of specific properties of supramolecular assemblies and nanomaterials so that hitherto inaccessible effects can be exploited for new applications. Examples are superparamagnetism or the high surface area helpful for catalysis and adsorption. In biology and medicine, superparamagnetic iron oxide nanoparticles have been used for cell selection and as magnetic resonance imaging (MRI) contrast agents. Furthermore, uptake of nanoparticles into a wide variety of cells is an effect that seems to be specific for materials in the range of 50-200 nm. Surface modifications (positively or negatively charged side groups of the polymers, amino acids, or peptides/proteins) enhance this uptake. Knowledge about factors influencing cellular uptake, like size, surface properties, cell type, and endocytotic pathways, enables optimization of labeling and selection of cells and nanoparticles for applications in vitro and in vivo. For in vivo applications, we will focus on how nanoparticles can cross the blood-brain barrier.
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