Reagents and Methods for Affinity Purifications
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9924 West 74th Street, Eden Prairie, MN, 55344
AbstractWe will develop reagents and methods for immobilizing receptors, including antibodies andStaphylococcal Protein A, onto membranes for affinity purifications, by photoimmobilizing hydrophilicpolymeric coupling reagents to which the receptors will then be coupled. Reagents and methods willincrease stability of the immobilized receptor or antibody molecules. This approach will synthesizehydrophilic polymers having photoactivatable groups and multiple groups that will react with antibodiesor other receptors. The polymeric coupling reagents will be designed to form very open polymeric layerswhen immobilized to allow penetration of the receptors into the layer to achieve efficient coupling whilealso having very low nonspecific adsorption of proteins in the solutions being purified. The technologywill be developed using model polyclonal antibodies after which it will also be Staphylococcus aureusProtein A. Use of these methods for preparing affinity membranes is expected to increase the efficiencyof the purification process, to improve the quality of such purified products and to reduce the costs ofa variety of bioseparations, including purification of recombinant proteins, antibodies and plasma proteinsfrom cell culture supernatants, plasma and ascites fluid.
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