The development of a new technology for reagentless multiplexed determination of protein concentrations in clinical and biological samples is proposed. The proposed approach employs nanotubes with molecular sized openings embedded within a mechanical and chemically robust polymeric membrane. We propose that a microarray may be constructed using these membranes in which isolated groups of nanotubes are fabricated as separate wells in the membrane, each group/well derivatized with a different molecular recognition agent (MRA), such as an antibody. In operation, an array would be spotted first in each well with MRAs to the proteins of interest, and then exposed to the analyte solution containing one or more of these protein analytes. The protein analytes in the mixture would form binding partners (e.g., antibody/antigen complex) with the immobilized MRAs. The array would then be """"""""read out"""""""" by a novel development and optical imaging process, to yield the protein concentration in the sample associated with each MRA. The arrays can be made in microscope slide format for compatibility with commercial microarray readers. In the proposed approach, proteins in a sample are detected directly, with no protein analyte premodification or sandwich formation with a labeled secondary antibody. Furthermore, the method is capable of single molecule sensitivity (if desired). The core concepts of this technology have already been developed and demonstrated by Prof. Charles Martin's research group at the University of Florida. This STTR effort will team Martin's group with EIC Laboratories to develop this new technology for proteomic microarrays. The 12 month Phase I study is designed to demonstrate the feasibility of the approach by establishing membrane production methods, demonstrating the underlying optical imaging phenomenon, establishing quantitative assay characteristics and coefficients of variation, and demonstrating optical imaging formats. Commercial EIC Laboratories will commercialize the technology by manufacturing and direct sales of the arrays as blanks and with selected MRAs. The market and products are similar to existing protein microarray products, with the new approach offering more rapid and sensitive testing. The proposed microarrays will find applications in disease detection/progression, analysis of protein expression levels, protein analysis for small samples such as biopsies, identification of protein-protein interactions for research, drug development, etc., biomarker discovery. These applications already guide the substantial proteomic microarray market; the proposed technology is expected to offer significant improvements in time, cost and sensitivity to existing approaches. ? ? ?