Immobilizing an Entity in a Desired Orientation on a Support Material
AssigneeArizona Board of Regents, acting for and on behalf of Arizona State University
InventorsStephen A. Johnston, Chris W. Diehnelt
Life of this patent
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Abstract
The present invention relates to the identification and selection of attachment molecules that attach/immobilize an entity having a detectable activity or property on a support in an orientation that provides a detectable activity or property, and to surfaces made of the attachment molecules.
Claims40
A method of identifying one or more attachment molecules for immobilizing an entity having an active region to a surface of a support material in an orientation that optimizes the availability of the active region for binding to a target, comprising
- providing a surface to which a plurality of different molecules is bound at respective different locations on the surface of the support material in a microarray;
- providing the entity,
- reacting the entity with the target to form an entity-target complex,
- contacting the plurality of different molecules with the entity-target complex under conditions that permit the entity-target complex to bind to the different molecules;
- detecting binding of the entity-ligand complex at each respective different location, and if binding is detected, and
- selecting the molecule or molecules for immobilizing the entity at those locations where the entity binds the target.
A method of identifying one or more attachment molecules for immobilizing an entity having an active region to a surface of a support material in an orientation that optimizes the availability of the active region for binding to a target, comprising
- providing a surface to which a plurality of different molecules is bound at respective different locations on the surface in a microarray;
- contacting the plurality of molecules with the entity under conditions that permit the entity to bind to the molecules;
- contacting the entity with the target under conditions that permit the entity to bind to the target,
- determining whether the entity binds the target at each respective different location;
- and
- selecting the molecule or molecules for immobilizing the entity at those locations where the entity binds the target.
The method as in one of claims 25-27 or claim 6, wherein the amino acid sequences of the plurality of different attachment peptides are generated by a pseudorandom computational process or are selected from a peptide library.
The surface of claim 25, wherein the entity is carbonic anhydrase and the target is dansylamide.
The surface of claim 25, where the entity is beta-galactosidase and the target is beta-galactoside.
The surface as in claim 29, wherein the sequences of the polypeptides are generated by a pseudorandom computational process.
The surface as in one of claims 25-27 wherein the attachment molecules are bound to the surface by a covalent bond.
The surface as in one of claims 25-27 wherein the attachment molecules are bound to the entity by a covalent bond.
An attachment peptide having the amino acid sequence FRNFPVPVIFRYLNPWPGSC, or a fragment or variant thereof.
An attachment peptide that is a member selected from the group consisting of CA4, CA8, and CA11.
A surface for immobilizing an entity having an active region, comprising a support material to which a plurality of attachment molecules bound thereby creating a surface of attachment molecules capable of binding and immobilizing the entity in an orientation that optimizes accessibility of the active region for binding to a target.
A surface for immobilizing an entity having an active region, comprising a support material to which a plurality of attachment molecules bound thereby creating a surface of attachment molecules capable of binding and immobilizing the entity in an orientation that reduces accessibility of the active region for binding to a target.
A surface for immobilizing an entity having a detectable property, comprising a support material to which a plurality of attachment molecules bound thereby creating a surface of attachment molecules capable of binding and immobilizing the entity in an orientation that optimizes the detectable property.
Dependencies are inferred from the claim text.