Passivation of surfaces after ligand coupling
Passivated substrates are provided for use in assays, comprising at least one covalently bonded ligand having specific binding activity for a molecule, and at least one covalently bonded blocking agent, wherein said ligand is directly bonded to the substrate surface. In certain embodiments, the ligand and blocking agent are covalently bonded only to the substrate surface, and not directly bonded to each other. In certain other embodiments, the ligand and blocking agent have at least one additional covalent bond to one another. Methods for preparing and using passivated substrates in bioassays are also provided.
1. A passivated substrate for use in assays, comprising
at least one covalently bonded ligand having specific binding activity for a molecule, wherein the covalently bonded ligand is directly bonded to a substrate; and
at least one covalently bonded blocking agent, wherein the blocking agent is a hydrophilic polymer directly bonded to the substrate; and, further the blocking agent and the ligand have at least one additional covalent bond to one another.
2. The passivated substrate of claim 1 , wherein the hydrophilic polymer is a polysaccharide, polypeptide or polyoxyethylene oxide.
3. The passivated substrate of claim 2 , wherein the polysaccharide is an aminosaccharide having a molecular weight of from 1 kDa to 500 kDa.
4. The passivated substrate of any of claims 1 , 2 - 3 , wherein the substrate is selected from a particle, a filter, a fiber, a glass slide, a microtiter plate and a microfluidic device.
5. The passivated substrate of claim 4 , wherein the substrate comprises a glass, a ferromagnetic material, or a polymer selected from a polystyrene, an acrylate, a methacrylate, or a combination or composite thereof.
6. The passivated substrate of claim 1 , wherein the ligand is a biopolymer selected from the group consisting of an antibody, a diabody, a receptor, an enzyme, a lectin, a peptide, a natural peptide, a saccharide, an oligonucleotide, a biotin binding ligand, and an immunoglobulin binding protein.
7. The passivated substrate of claim 6 , wherein the ligand has a binding affinity of at least 10 6 M −1 for the molecule.
8. The passivated substrate of claim 3 , wherein the polysaccharide is an aminosaccharide having a molecular weight of from 10 kDa to 40 kDa.