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| - Immunoassay Reagents for Thyroid Testing. 3. Determination ofthe Solution Binding Affinities of a T4 Monoclonal Antibody FabFragment for a Library of Thyroxine Analogs Using SurfacePlasmon Resonance
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| - A library of thyroxine analogs and tracers was prepared, and theirsolution binding affinities for ananti-T4 Fab fragment were determined using a singlehigh-density l-T4 biosensor surface in aBIAcoresurface plasmon resonance instrument. The high-densityl-T4 analog biosensor was calibratedbydetermination of the initial binding rate was of known concentrationsof free anti-T4 Fab fragment insolution to the biosensor surface. A range of individual thyroxineanalog and tracer concentrationswas subsequently mixed with a fixed concentration ofanti-T4 Fab fragment. The concentrationoffree anti-T4 Fab fragment in each solution at equilibriumwas determined, and the equilibriumdissociation constant (KD) for each case wasderived. The KD values determined insolution arecompared to values determined by a direct kinetic analysis on theBIAcore instrument using individualbiosensor surfaces.
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