Entrapped binding protein as biosensors

A method of making a biosensor, said biosensor comprising a crosslinked polymeric hydrogel and a binding molecule, said binding molecule being covalently attached to said hydrogel and capable of generating a detectable signal directly upon binding of a target molecule to said binding molecule, said method comprising polymerizing and crosslinking monomers in the presence of water and said binding molecule.

A method of detecting an analyte in a sample comprising

• a) providing a biosensor comprising a crosslinked, polymeric hydrogel and a binding molecule, said binding molecule being covalently attached to said hydrogel and being capable of generating a detectable signal directly upon binding of said analyte to said binding molecule
• b) contacting said biosensor with said sample
• c) comparing the signal generated by said binding molecule when said biosensor is contacted with said sample with the signal generated by said binding molecule when said biosensor is contacted with an analyte-free control sample, wherein a difference in the signal generated by said binding molecule when said biosensor is contacted with said test sample, as compared to when said biosensor is contacted with said control sample, indicates that test sample contains the analyte.

A method of making a polymeric hydrogel biosensor, said method comprising polymerizing a functionally derivatized binding protein and at least one monomer to produce said crosslinked polymeric hydrogel biosensor

A method of making a crosslinked polymeric hydrogel biosensor, said method comprising photo polymerizing a monomer in the presence of a binding protein to produce said polymeric hydrogel biosensor comprising said binding protein.

A biosensor comprising a binding protein wherein said biosensor has an apparent dissociation constant (Kd) of at least one order of magnitude greater than the Kd of said free binding protein in solution.