Entrapped binding protein as biosensors
First Claim
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1. A glucose biosensor for in vivo or in vitro use comprising:
- a) at least one mutant glucose/galactose binding protein (GGBP) and at least one reporter group covalently attached thereto wherein said mutant GGBP comprises at least one amino acid substitution of a non-reactive amino acid with a reactive amino acid, wherein said at least one amino acid substitution is selected from the group consisting of a cysteine at position 11, a cysteine at position 14, a cysteine at position 19, a cysteine at position 43, a cysteine at position 74, a cysteine at position 107, a cysteine at position 110, a cysteine at position 112, a cysteine at position 113, a cysteine at position 137, a cysteine at position 149, a cysteine at position 213, a cysteine at position 216, a cysteine at position 238, a cysteine at position 287, a cysteine at position 292, a cysteine at position 112 and a serine at position 238, a cysteine at position 149 and a serine at position 238, a cysteine at position 152 and a cysteine at position 182, a cysteine at position 152 and a serine at position 213, a cysteine at position 213 and a cysteine at position 238, a cysteine at position 149 and an arginine at position 213, a cysteine at position 149 and a serine at position 213 and a serine at position 238, and a cysteine at position 149 and an arginine at position 213 and a serine at position 238 corresponding to amino acid positions of E. coli GGBP, andb) a first analyte permeable matrix entrapping or encapsulating said at least one mutant GGBP, said matrix comprising a covalently crossliniked hydrogel or a sol-gel comprising organically modified silicates or a combination thereof, wherein the first analyte permeable matrix is formulated to allow three-dimensional conformational movement of the at least one mutant GGBP to bind and release the glucose, and wherein the matrix is adapted to allow glucose to diffuse into and out of the matrix.
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Abstract
The invention is directed to entrapped mutated binding proteins, mutated binding proteins containing reporter groups, compositions of mutated binding proteins containing reporter groups in analyte permeable matrixes, and their use as analyte biosensors both in vitro and in vivo.
63 Citations
28 Claims
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1. A glucose biosensor for in vivo or in vitro use comprising:
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a) at least one mutant glucose/galactose binding protein (GGBP) and at least one reporter group covalently attached thereto wherein said mutant GGBP comprises at least one amino acid substitution of a non-reactive amino acid with a reactive amino acid, wherein said at least one amino acid substitution is selected from the group consisting of a cysteine at position 11, a cysteine at position 14, a cysteine at position 19, a cysteine at position 43, a cysteine at position 74, a cysteine at position 107, a cysteine at position 110, a cysteine at position 112, a cysteine at position 113, a cysteine at position 137, a cysteine at position 149, a cysteine at position 213, a cysteine at position 216, a cysteine at position 238, a cysteine at position 287, a cysteine at position 292, a cysteine at position 112 and a serine at position 238, a cysteine at position 149 and a serine at position 238, a cysteine at position 152 and a cysteine at position 182, a cysteine at position 152 and a serine at position 213, a cysteine at position 213 and a cysteine at position 238, a cysteine at position 149 and an arginine at position 213, a cysteine at position 149 and a serine at position 213 and a serine at position 238, and a cysteine at position 149 and an arginine at position 213 and a serine at position 238 corresponding to amino acid positions of E. coli GGBP, and b) a first analyte permeable matrix entrapping or encapsulating said at least one mutant GGBP, said matrix comprising a covalently crossliniked hydrogel or a sol-gel comprising organically modified silicates or a combination thereof, wherein the first analyte permeable matrix is formulated to allow three-dimensional conformational movement of the at least one mutant GGBP to bind and release the glucose, and wherein the matrix is adapted to allow glucose to diffuse into and out of the matrix. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A glucose biosensor for in vivo or in vitro use comprising:
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a) at least one mutant glucose/galactose binding protein (GGBP) and at least one reporter group covalently attached thereto wherein said mutant GGBP comprises at least one amino acid substitution of a non-reactive amino acid with a reactive amino acid, and b) a first analyte permeable matrix entrapping or encapsulating said at least one mutant GGBP, said matrix comprising a covalently crossliniked sol-gel comprising organically modified silicates, wherein said sol-gel is selected from at least partially cured hydrolytically condensable siloxanes condensed with at least one water soluble organic polyol component, wherein the first analyte permeable matrix is formulated to allow three-dimensional conformational movement of the at least one mutant GGBP to bind and release the glucose, and wherein the matrix is adapted to allow glucose to diffuse into and out of the matrix. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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Specification