PROTEOMICS · 2004 · 154 citations · 18 references
Life SciencesBiotechnologyMolecular BiologyNanobiotechnologyProtein-protein InteractionsProteomicsAtomic Force MicroscopyFluorescence MicroscopyMicrofluidicsMolecular ScreeningProtein MicroarraysAntibodiesFunctional AssaysMicroarray AnalysisMicrofluidic PlatformsMultiplex Microarray AssaysMultiplexed AssaysProtein-protein Interaction DetectionSample Size Reduction
The use of microarrays for parallel screening of nucleic acid profiles has become an industry standard. Similar efforts for screening protein-protein interactions are gaining momentum, however, they remain limited by the requirement for relatively large sample volumes. One strategy for overcoming this problem is to significantly decrease the size and consequently the sample volume of the protein interaction assay. We report here on our progress over the last two years in the construction of ultraminiaturized, functional protein capture assays. Each one micron spot in these array-based assays covers less than 1/1000(th) of the surface area of a conventional microarray spot while still maintaining enough antibodies to provide a useful dynamic range. These nanoarray assays can be read by conventional optical fluorescence microscopy as well as by novel label-free methods such as atomic force microscopy. The size reduction realized by functional protein nanoarrays also creates opportunities for novel applications including highly multiplexed single cell analysis and integration with microfluidics and other "lab-on-a-chip" technologies.
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