초록 ▼

Methods for preparing a biochip are provided herein wherein the biomolecular probe to be used with the biochip is alternatively bound to a hydrogel prepolymer prior to or simultaneously with polymerization of the prepolymer. In particularly preferred embodiments, a polyurethane-based hydrogel prepol

Methods for preparing a biochip are provided herein wherein the biomolecular probe to be used with the biochip is alternatively bound to a hydrogel prepolymer prior to or simultaneously with polymerization of the prepolymer. In particularly preferred embodiments, a polyurethane-based hydrogel prepolymer is derivatized with an organic solvent soluble biomolecule, such as a peptide nucleic acid probe in aprotic, organic solvent. Following derivatization of the prepolymer, an aqueous solution, for example sodium bicarbonate, preferably buffered to a pH of about 7.2 to about 9.5, is added to the derivatized prepolymer solution to initiate polymerization of the hydrogel. Alternatively, a water soluble biomolecule, such as DNA or other oligonucleotide, is prepared in an aqueous solution and added to the polyurethane-based hydrogel prepolymer such that derivatization and polymerization occur, essentially, simultaneously. While the hydrogel is polymerizing, it is microspotted onto a solid substrate, preferably a silanated glass substrate, to which the hydrogel microdroplet becomes covalently bound. Most preferably the hydrogel microdroplets are at least about 30 .mu.m thick, for example about 50 .mu.m to about 100 .mu.m thick. The resulting biochips are particularly useful for gene discovery, gene characterization, functional gene analysis and related studies.

대표청구항 ▼

[ What is claimed is:] [1.] A method of preparing a biochip having an organic solvent soluble biomolecule covalently bound thereto, the method comprising:a) providing a solid substrate having a top surface with reactive molecules thereon;b) providing a solution comprising the biomolecule in an aprot

[ What is claimed is:] [1.] A method of preparing a biochip having an organic solvent soluble biomolecule covalently bound thereto, the method comprising:a) providing a solid substrate having a top surface with reactive molecules thereon;b) providing a solution comprising the biomolecule in an aprotic organic solvent;c) providing a polyurethane-based hydrogel prepolymer in an aprotic organic solvent;d) derivatizing the polyurethane-based hydrogel prepolymer with the biomolecule from step (b);e) initiating polymerization of the derivatized polyurethane-based hydrogel prepolymer of step (d) by adding a buffered aqueous solution thereto; andf) microspotting the polymerizing hydrogel solution of step (e) onto the top surface of the substrate of step (a), such that the polymerizing hydrogel reacts with the reactive molecules on the surface of the substrate resulting in covalent binding of the hydrogel to the substrate.