Fabrication and Imaging of Protein Crossover Structures
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Fabrication and Imaging of Protein Crossover Structures John R. LaGraff,1 Yi-Ping Zhao,2 David J. Graber,3 Dan Rainville,4 Gwo-Ching Wang,5 TohMing Lu,5 Quynh Chu-LaGraff,6 Don Szarowski,3 William Shain,3 James N. Turner3 1 Department of Chemistry, Hamilton College, Clinton, NY 13323, USA 2 Department of Physics and Astronomy, University of Georgia, Athens, GA 30602, USA 3 Wadsworth Center for Laboratory Research, New York State Department of Health, Albany, NY 12201, USA 4 Department of Physics, Siena College, Loudonville, NY 12211, USA 5 Department of Physics, Applied Physics and Astronomy, Rensselaer Polytechnic Institute, Troy, N.Y, 12180 USA 6 Department of Biology, Union College, Schenectady, NY 12308, USA
ABSTRACT Proteins often deform, dehydrate or otherwise denature when adsorbed or patterned directly onto an inorganic substrate, thus losing specificity and biofunctionality. One method used to maintain function is to pattern the protein of interest directly onto another underlying protein or polypeptide that acts as a buffer layer between the substrate and the desired protein. We have used microcontact printing (µcp) to cross-stamp orthogonal linear arrays of two different proteins (e.g., IgG, poly-lysine, protein A) onto glass substrates. This created three separate types of protein-substrate microenvironments, including crossover structures of protein one on protein two. We report preliminary fluorescent microscopy and scanning force microscopy characterization of these structures, including commonly encountered structural defects.
INTRODUCTION Mono-molecular layers of proteins patterned onto surfaces at the micron and nanoscale can potentially serve in a number of useful capacities such as active components of biosensors [1], regulators of directed cell growth [2], and diagnostic microarrays or protein chips [3]. For example, protein chips are being developed in an effort to miniaturize biological assays by placing patterned arrays of multiple types of proteins onto surfaces [3]. Such protein chips will allow simultaneous detection and analysis of multiple species, require smaller quantities of expensive reagents, and have more rapid biochemical reactions due to short mass transport distances [3]. Biomedical industry will also benefit from improved design and patterning of protein chips for high-throughput detection and profiling of various interactions such as drugprotein, protein-protein, and disease antigen-antibody [3]. Patterning a chip containing hundreds if not thousands of different proteins—each with its own distinctive protein-substrate interaction forces—and having all proteins retain their unique three-dimensional shape and biological function is a formidable challenge. Although substantial investments are being poured into the commercial development of protein chips, reliability and reproducibility remain elusive owing primarily to an incomplete understanding and control of fundamental proteinsubstrate interactions during patterning.
Mat. Res. Soc. Symp. Proc. Vol. 735 © 2003 Mat
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