Streptavidin properties

While a number of proteins have been crystallized in this manner, the majority of studies have focused on a robust system comprising the tetrameric protein streptavidin and the vitamin biotin. The choice of this system is primcirily motivated by the strong bond between biotin and streptavidin (having an association equilibrium constant, Ka Tbe binding properties were recently... 

Agiamamioti and co-workers [72] synthesized a novel biotinylated fluorophore, 10-(2-biotinyloxyethyl)-9-acridone 25 with favorable properties for bioanalytical applications. In aqueous solutions, it displayed high fluorescence ([Pg.37]

Luschtinetz F, Dosche C, Kumke MU (2009) Influence of streptavidin on the absorption and fluorescence properties of cyanine dyes. Bioconjugate Chem 20 576-582... 

D protein arrays based on biotin-streptavidin architectures are likely to be the system of choice due to their ease in handling, excellent signal-to-noise ratio and non-specific interactions. 3D surfaces based on porous gold, sol-gel materials, polymer brushes and dextran surfaces are widely used to mimic the properties of bulk solution and increase the immobilization capacity of proteins. 

The spectral properties of four major phycobiliproteins used as fluorescent labels can be found in Tables 9.1 and 9.2. The bilin content of these proteins ranges from a low of four prosthetic groups in C-phycocyanin to the 34 groups of B- and R-phycoerythrin. Phycoerythrin derivatives, therefore, can be used to create the most intensely fluorescent probes possible using these proteins. The fluorescent yield of the most luminescent phycobiliprotein molecule is equivalent to about 30 fluoresceins or 100 rhodamine molecules. Streptavidin-phycoerythrin conjugates, for example, have been used to detect as little as 100 biotinylated antibodies bound to receptor proteins per cell (Zola et al., 1990). 

Biotin modification reagents are widely used to attach a biotin group to proteins or other molecules for subsequent use in avidin, streptavidin, or NeutrAvidin separations or assays. Traditional biotin compounds containing aliphatic or other hydrophobic linker arms are discussed in detail in Chapter 11. In this section, the biotin-PEG compounds exclusively are discussed due to their unique hydrophilic properties, which include low nonspecific binding character and low immunogenicity. 

Chaiet, L., and Wolf, F.J. (1964) The properties of streptavidin, a biotin-binding protein produced by Streptomycetes. Arch. Biochem. Biophys. 106, 1-5. 

Guided mode calculations were also carried out to compare the sensor response of several waveguide systems. In these simulations a model molecular monolayer is represented by a 2-nm thick layer with a refractive index of n 1.5. The optical properties of this model layer are typical of a dense layer of organic molecules on a substrate1 41, and are a reasonable approximation for a streptavidin protein layer bound to a biotinylated surface, the experimental model system we use to characterize our sensors. The ambient upper cladding was assumed to be water with a refractive index of n 1.32. For all examples, the lower cladding was assumed to be Si02 with an index of n 1.44. In the simulations, the effective index of... 

X-streptavidin. The intricate interplay between the steric and electronic properties of the acceptor and the polymeric donor may have important impact for the design of future biosensors. 

Because of concerns about the safety of radioisotope use, researchers are developing fluorescent and chemiluminescent methods for detection of small amounts of biomolecules on gels. One attractive approach is to label biomolecules before analysis with the coenzyme biotin. Biotin forms a strong complex with enzyme-linked streptavidin. Some dynamic property of the enzyme is then measured to locate the biotin-labeled biomolecule on the gel. These new methods approach the sensitivity of methods involving radiolabeled molecules, and rapid advances are being made. 

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