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US 5,766,908 AGranted
High-flux semipermeable membrane containing immobilized affinity ligands
AssigneeAkzo Nobel N.V.
InventorsElias Klein, Donald H. Yeager
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AssigneeAkzo Nobel N.V.
InventorsElias Klein, Donald H. Yeager
Estimating the term…
An affinity support is provided containing a high flux semipermeable hydrogel membrane surface-modified with an affinity ligand, such as a protein which may be an antibody or enzyme, or a cell receptor complement, useful for affinity separation of biological macromolecules, including insoluble proteins, cells, and cell fragments, from solution. The exclusion limit (molecular weight cut-off) of the matrix is selected to substantially restrict immobilized protein or other ligand to the surface thereof for maximization of available ligand binding capacity. The exclusion limit is also selected to permit reagent(s) used for the matrix/ligand linkage to penetrate into and form covalent bonds with the membrane on the interior surfaces of the membrane for optimizing packing densities of the affinity ligand exterior to the membrane. By this substantially complete orientation of affinity ligand between affinity matrix and solution interface, the recognition sites of the proteins or other ligands are maximally available to complementary molecules or cells bearing complementary determinants for complexing therewith.
An affinity support for affinity separation including an affinity matrix comprising a high-flux semipermeable hydrogel membrane surface-modified with one or more immobilized affinity ligands, said membrane having free residual functional groups for immobilization of the affinity ligand and pores of a size to provide a nominal molecular weight cutoff small enough to essentially restrict the immobilized ligand outwardly of the exterior surface of the matrix and large enough to permit entry of an immobilizing reagent for ligand immobilization into the interior pores of the membrane for coupling therein, and said immobilized affinity ligand being outwardly spaced from the exterior surface of the membrane and indirectly coupled to a functional group within a pore thereof via the immobilizing reagent wherein one end of the reagent is coupled to the functional group within a pore of the membrane and the other end thereof extends outwardly of the exterior surface of the membrane and is coupled to the ligand.
Dependencies are inferred from the claim text.