Stabilization of activated galactose oxidase enzyme
First Claim
1. In a method for the quantitative determination of a substance which reacts with a galactose oxidase enzyme in a manner whereby a reactant or a product of said reaction may be analytically measured and wherein said substance is introduced for determination by being included in a buffer solution which is brought into contact with said enzyme, said buffer solution also containing a sufficient amount of a reduction-oxidation agent to activate said enzyme, the improvement comprising adding to the buffer solution which contacts said galactose oxidase enzyme a sufficient amount of cupric ion to stabilize said activated enzyme whereby the stabilized, activated galactose oxidase enzyme may be reused and stored for a period of time after use without loss of its enzymic activity.
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Abstract
Galactose oxidase enzyme can effectively be used for the determination of a number of different substances if activated and stabilized in accordance with the present invention. Activation is accomplished with a redox agent such as ferricyanide. Stabilization of the activated galactose oxidase is accomplished by addition of cupric ion. Upon stabilization, activated, immobilized galactose oxidase enzymes may be stored and reused for at least 25 days.
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Citations
13 Claims
- 1. In a method for the quantitative determination of a substance which reacts with a galactose oxidase enzyme in a manner whereby a reactant or a product of said reaction may be analytically measured and wherein said substance is introduced for determination by being included in a buffer solution which is brought into contact with said enzyme, said buffer solution also containing a sufficient amount of a reduction-oxidation agent to activate said enzyme, the improvement comprising adding to the buffer solution which contacts said galactose oxidase enzyme a sufficient amount of cupric ion to stabilize said activated enzyme whereby the stabilized, activated galactose oxidase enzyme may be reused and stored for a period of time after use without loss of its enzymic activity.
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6. A method for the quantitative polarographic determination of a polarographically inactive material which reacts with galactose oxidase enzyme in a manner whereby a reactant or a product of said reaction may be polarographically measured, comprising:
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(a) providing a polarographic cell including at least one electrode positioned behind a membrane permeable to the material being measured and in contact with an electrolyte, (b) establishing a potential across said cell such that a current is produced which is proportional to the amount of hydrogen peroxide or oxygen present on the electrode side of said membrane, (c) bringing said cell into contact with a quantity of material containing said polarographically inactive material in the presence of galactose oxidase enzyme and a buffer containing a reduction-oxidation agent said reduction-oxidation agent being present in said buffer in a sufficient amount to activate the galactose-oxidase enzyme, (d) adding to said buffer a sufficient amount of cupric ion to stabilize the activated enzyme, and (e) determining the current flowing across said cell as an indication of the amount of said substance present in said quantity of material. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13)
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Specification