Immobilization of active agents on substrates with a silane and heterobifunctional crosslinking agent
First Claim
1. A method for modifying a substrate to activate said substrate for the immobilization of anzime group-containing active agents, said method comprising,selecting a substrate having hydroxyl groups on at least one surface,coating said hydroxyl-containing surface with a thiol-terminal group silane containing a reactive group other than a thiol group that reacts only with the hydroxyl groups on said surface to form a silane layer having an available reactive thiol group,coating said silane layer with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts only with the thiol-terminal group on said silane to form a layer of crosslinking agent having an available free succinimide reactive group capable of reacting with amino group of said active agent.
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Accused Products
Abstract
Active agents such as proteins are covalently immobilized on substrates carrying hydroxyl groups. A silane is bound to the substrate and coupled to a heterobifunctional crosslinker at one functional group leaving a free functional group, different than the first group, to which a protein is bound while retaining high protein functionality. Preferably, the silane has a functional group which reacts with the hydroxyl group of the substrate and a thiol terminal group which reacts with a functional group of a heterobifunctional crosslinking agent which contains a succinimide group that reacts with an amino group of the active agent. Bound active agents such as proteins are useful as biosensors or reactants in a variety of applications including bioassays.
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Citations
27 Claims
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1. A method for modifying a substrate to activate said substrate for the immobilization of anzime group-containing active agents, said method comprising,
selecting a substrate having hydroxyl groups on at least one surface, coating said hydroxyl-containing surface with a thiol-terminal group silane containing a reactive group other than a thiol group that reacts only with the hydroxyl groups on said surface to form a silane layer having an available reactive thiol group, coating said silane layer with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts only with the thiol-terminal group on said silane to form a layer of crosslinking agent having an available free succinimide reactive group capable of reacting with amino group of said active agent.
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22. In a method of immobilizing an amino group-containing active agent on a substrate, the improvement comprising preparing said substrate by,
selecting a substrate having hydroxyl groups on a surface thereof, depositing on the substrate of film of a thiol-terminal group silane having a reactive group other than the thiol group that reacts with said hydroxyl groups to form silane surface having a free reactive thiol group, coating said silane surface with a heterobifunctional succinimide group-containing crosslinking agent having a functional group other than the succinimide group that reacts with said free reactive thiol group to form a layer of the crosslinking agent having an available free succinimide reactive group that reacts with the amino group of said active agent.
Specification