Methods of immobilizing ligands on solid supports and apparatus and methods of use therefor
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.
Claims58
A method of immobilizing an affinity ligand on a solid support comprising:
- providing a solid support comprising an immobilized thiol group;
- contacting the thiol group with a nucleic acid comprising an acrylamide functional group;
- and
- forming a covalent bond between the two groups,
- thereby immobilizing the ligand on the solid support.
A method of immobilizing an affinity ligand on a solid support comprising the steps of:
- providing a solid support comprising immobilized latent thiol groups;
- activating the latent thiol groups;
- and
- reacting the activated thiol groups with an affinity ligand having at least one acrylamide functional group, thereby immobilizing an affinity ligand on a solid support.
The method of claim 26, wherein R4 is selected from the group consisting of:
- 30
- wherein R8 is a substituted or unsubstituted aliphatic group, a substituted or unsubstituted aromatic group, a substituted or unsubstituted heteroaromatic group, a substituted or unsubstituted aralkyl, or a substituted or unsubstituted heteroaralkyl group.
A method of forming an array of nucleic acids immobilized on a solid support comprising:
- forming an amine-derivatized region on the support;
- treating the amine-derivatized region with a thiolating agent such that latent thiol groups immobilized on the support are formed;
- activating the latent thiol groups;
- contacting the activated thiol groups with a plurality of nucleic acids comprising acrylamide functional groups;
- and
- forming a covalent bond between the two groups, thereby forming an array of nucleic acids immobilized on the solid support.
A method of preparing a solid support having immobilized thiol groups comprising contacting a solid support with a silane compound represented by the formula
- RnSiX(4−n)
- wherein X is selected from the group consisting of a halogen, an alkoxide, an alkane, an acyloxy, and an amine, and R is a compound having at least one substituent selected from the group consisting of an a,P-unsaturated carbonyl and a vinyl.
Dependencies are inferred from the claim text.