Assay for neuraminic acids
First Claim
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1. A composition having a pH of between 5 and 9 comprising at least 0.1 unit neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.2 millimolar to about 10 millimolar electron acceptor.
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Abstract
A composition and element for assaying a neuraminic acid comprises a neuraminic acid aldolase, pyruvate oxidase and an electron acceptor. A sample containing a neuraminic acid can be contacted with the composition and the product detected spectrophotometrically.
9 Citations
33 Claims
- 1. A composition having a pH of between 5 and 9 comprising at least 0.1 unit neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.2 millimolar to about 10 millimolar electron acceptor.
- 7. An aqueous composition having a pH of between 5 and 9 comprising a liquid containing a neuraminic acid, at least 0.1 unit neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.02 millimolar to about 10 millimolar of an electron acceptor.
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13. An aqueous composition having a pH between about 5 and 9 comprising a biological sample containing sialic acid, at least 0.1 unit of N-acetyl neuraminic acid aldolase, at least about 0.005 unit of pyruvate oxidase, from about 0.02 millimolar to about 10 millimolar of a dichloroindophenol and at least about 0.01 unit of neuraminidase.
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14. A method for the detection of a neuraminic acid comprising the steps of:
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(a) contacting in an aqueous medium a sample for analysis and a composition having a pH between about 5 and 9 comprising at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.02 millimolar to about 10 millimolar of an electron acceptor; and (b) determining the detectable product. - View Dependent Claims (15, 16, 17, 18, 25)
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19. A method for the detection of a neuraminic acid bound to other biological materials comprising the steps of:
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(a) contacting in an aqueous medium a sample for analysis and a composition having a pH between about 5 and 9 comprising at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase, between about 0.02 millimolar to about 10 millimolar of an electron acceptor and at least 0.01 unit neuraminidase; and (b) determining the detectable product.
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20. A method for the detection of free and bound neuraminic acids comprising the steps of:
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(a) contacting in an aqueous medium a sample for analysis and a composition having a pH between about 5 and 9 comprising at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.02 millimolar to about 10 millimolar of an electron acceptor; (b) determining any detectable product of (a); (c) adding to said aqueous medium at least 0.01 unit neuraminidase; and (d) determining the detectable product of (c); and (e) quantifying the amount of free neuraminic acid and bound neuraminic acid from the difference of (b) and (d). - View Dependent Claims (21, 22, 23)
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- 24. An element for the detection of a neuraminic acid comprising a support having thereon at least one reagent layer containing at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.02 millimolar to about 10 millimolar of an electron acceptor.
- 29. An element for the detection of neuraminic acids bound to other biological materials comprising a support having thereon a composition having a pH between about 5 and 9 comprising at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase, from about 0.02 millimolar to about 10 millimolar of an electron acceptor and at least 0.01 unit neuraminidase.
- 32. An element for the detection of a neuraminic acid in a liquid, the element comprising a support having thereon a spreading layer, at least one reagent layer and a registration layer in fluid contact under conditions of use, the reagent layer intervening the spreading layer and the registration layer, and the registration layer intervening the reagent layer and the support, the reagent layer comprising at least 0.1 unit of a neuraminic acid aldolase, at least about 0.005 unit pyruvate oxidase and from about 0.2 millimolar to about 10 millimolar of an electron acceptor.
Specification