Method of forming a molded protein matrix for a sensor
First Claim
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1. A method of forming a molded protein matrix for a sensor comprising the steps of:
- combining a protein mixture with a first cross-linking reagent;
placing the combined protein mixture and first cross-linking reagent into a mold, where the mold comprises an internal volume;
incubating the combined protein mixture and first cross-linking reagent in the internal volume of the mold for a sufficient time to form a molded protein matrix having a three-dimensional shape that corresponds to the internal volume of the mold;
after incubating, exposing the molded protein matrix in the mold to a second cross-linking reagent for a time sufficient to cause the molded protein matrix to become further hardened than after the incubating, resulting in the formation of a hardened molded protein matrix.
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Abstract
A method for formulating and immobilizing a protein and a protein matrix formed by the method. The protein matrix preparation method results in a physically and chemically stable protein matrix that has low swelling, non-leaching, high activity, and high mechanical strength properties. The method includes cross-linking and hardening the protein mixture and using a mold to form a protein into a desired shape and size.
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Citations
20 Claims
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1. A method of forming a molded protein matrix for a sensor comprising the steps of:
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combining a protein mixture with a first cross-linking reagent; placing the combined protein mixture and first cross-linking reagent into a mold, where the mold comprises an internal volume; incubating the combined protein mixture and first cross-linking reagent in the internal volume of the mold for a sufficient time to form a molded protein matrix having a three-dimensional shape that corresponds to the internal volume of the mold; after incubating, exposing the molded protein matrix in the mold to a second cross-linking reagent for a time sufficient to cause the molded protein matrix to become further hardened than after the incubating, resulting in the formation of a hardened molded protein matrix. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A method of forming a molded protein matrix for a sensor comprising the steps of:
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combining a protein mixture with a first cross-linking reagent to form a protein paste, wherein the protein mixture includes human serum albumin (HSA) in a concentration of between about 23-32.5% (w/v) and glucose oxidase; placing the protein paste into a mold, where the mold comprises an internal volume; incubating the protein paste in the internal volume of the mold for a sufficient time to form a molded protein matrix having a three-dimensional shape that corresponds to the internal volume of the mold; after incubating, exposing the molded protein matrix in the mold to a second cross-linking reagent for a time sufficient to cause the molded protein matrix to become further hardened than after the incubating such that a hardened molded protein matrix is formed in a hardened state that minimizes swelling that would deform a shape of a body of a sensor, wherein the sensor body has a space for receiving the hardened molded protein matrix, and such that the hardened molded protein matrix minimizes shrinkage that would create voids between the hardened molded protein matrix and the body of the sensor once the hardened molded protein matrix has been received within space of the body, wherein the second cross-linking reagent is a vaporized cross-linking agent; removing the hardened molded protein matrix from the mold; placing the hardened molded protein matrix into a cross-linking solution including a third cross-linking agent; removing the hardened molded protein matrix from the cross-linking solution; washing the hardened molded protein matrix in a washing solution selected from the group consisting of sterile deionized water, phosphate buffered saline, and glycine buffer; and placing the washed and hardened molded protein matrix into the space in the body of the sensor.
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20. A method of forming a molded protein matrix for a sensor comprising the steps of:
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combining a protein mixture with a first cross-linking reagent to form a protein paste; placing the protein paste into a mold, where the mold comprises an internal volume; incubating the protein paste in the internal volume of the mold for a sufficient time to form a molded protein matrix having a three-dimensional shape that corresponds to the internal volume of the mold; after incubating, exposing the molded protein matrix to a second cross-linking reagent; wherein the exposing to a second cross-linking reagent is for a time sufficient to harden the molded protein matrix such that the hardened molded protein matrix minimizes swelling that would deform a shape of a body of a sensor, wherein the sensor body has a space for receiving the molded protein matrix, and such that the hardened molded protein matrix minimizes shrinkage that would create voids between the molded protein matrix and the body of the sensor once the molded protein matrix has been received within the space of the sensor body; and placing the hardened molded protein matrix into the space in the body of the sensor.
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Specification