Methods of immobilizing ligands on solid supports
AssigneeMatrix Technologies Corporation
InventorsEzra S. Abrams, Tianhong Zhang, Slawomir Mielewczyk, Brian C. Patterson
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Abstract
A method is provided for immobilizing a ligand, e.g., a nucleic acid, on a solid support. The method includes providing a solid support containing an immobilized latent thiol group, activating the thiol group, contacting the activated thiol group with a nucleic acid comprising an acrylamide functional group, and forming a covalent bond between the two groups, thereby immobilizing the nucleic acid to the solid support. Kits containing the solid supports and method of utilizing the solid supports are also provided.
Claims9
A method of immobilizing an affinity ligand on a microarray comprising the steps of:
- providing a solid support comprising immobilized latent thiol groups by contacting an unsaturated aliphatic surface of the support with a polymerization solution and a polymerizable disulfide compound to form a polymer matrix;
- activating the latent thiol groups by contacting the matrix with a disulfide reducing agent;
- and
- reacting the activated thiol groups with an affinity ligand having at least one α,β unsaturated carbonyl functional group to thereby immobilize the affinity ligand on a microarray.
A method of preparing a solid support having immobilized thiol groups, comprising the steps of:
- contacting a glass solid support with a silane compound represented by the following structural formula:
- wherein:
- X is a halogen;
- and
- R1, R2, and R3, are selected from the group consisting of a halogen, an alkyl group, an alkenyl group or a group having at least one α,β-unsaturated carbonyl, provided that at least one of R1, R2, or R3 is an alkenyl group or a group having at least one α,β-unsaturated carbonyl, thereby forming a solid support having an unsaturated aliphatic surface;
- thereafter contacting the unsaturated aliphatic surface of the solid support with a polymerization solution containing a free radical initiator and disulfide bisacrylamide represented by the following structural formula:
- wherein:
- n and m are each, independently, a positive integer, thereby forming a solid support comprising immobilized latent thiol groups;
- and
- thereafter contacting the latent thiol groups with a disulfide reducing agent, thereby forming a solid support having immobilized thiol groups.
Dependencies are inferred from the claim text.