Microcontact printing of biotin for selective immobilization of streptavidin-fused proteins and SPR analysis
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In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (μCP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene ofStreptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of μμCP and cSA-fused proteins, is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications, one such being the use in protein-protein assays.
Keywordsmicrocontact printing (μCP) pattern generation protein-protein assay surface plasmon resonance
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- Dewez, J. L., J. B. Lhoest, E. Detrait, V. Berger, C. C. Dupont-Gillain, L. M. Vincent, Y. J. Schneider, P. Bertrand, and P. G. Rouxhet (1998) Adhesion of mammalian cells to polymer surfaces: from physical chemistry of surfaces to selective adhesion on defined patterns.Biomaterials 19: 1441–1445.CrossRefGoogle Scholar
- Sambrook, J. and D. Russell (2001)Molecular Cloning: A Laboratory Manual. 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar