Immunoglobulin Surface Adsorption Studied by Total Internal Reflection with Fluorescence Correlation Spectroscopy
Total internal reflection with fluorescence correlation spectroscopy (TIR/FCS) is a new technique for studying binding and surface diffusion of fluorescent molecules in equilibrium at a surface. We apply TIR/FCS to the rapidly reversible nonspecific adsorption of rhodamine-labeled immunoglobulin to fused silica coated with bovine serum albumin. The measured characteristic time is 6.3ms and is limited by the rate of diffusion in solution. TIR/FCS is shown to be a feasible technique for measuring nonspecific binding kinetic rates at equilibrium and should prove useful for measuring specific solute/surface site kinetic rates in systems where nonspecific binding is low.
KeywordsFuse Silica Bulk Concentration Total Internal Reflection Fluorescence Correlation Spectroscopy Fluorescent Molecule
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