Assay
First Claim
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1. An assay method for determining of the level of glycated hemoglobin within a sample, comprising:
- providing a sample comprising hemoglobin;
denaturing the hemoglobin to effect a conformational change which causes exposure of a zinc binding site;
capturing the hemoglobin molecules using zinc coupled to a support medium; and
determining the amount of glycated hemoglobin bound by the coupled zinc.
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Abstract
Compositions and assay methods for and relating to the detection of glycated hemoglobin are provided. In particular, the presence and level of glycated hemoglobin can be detected in a blood sample. The compositions comprise microparticles which have zinc coating for use in the improved binding of proteins exhibiting a zinc binding domain, in particular glycated hemoglobin.
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Citations
31 Claims
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1. An assay method for determining of the level of glycated hemoglobin within a sample, comprising:
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providing a sample comprising hemoglobin;
denaturing the hemoglobin to effect a conformational change which causes exposure of a zinc binding site;
capturing the hemoglobin molecules using zinc coupled to a support medium; and
determining the amount of glycated hemoglobin bound by the coupled zinc. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 31)
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16. An assay method for the determination of the level of glycated hemoglobin within a sample, comprising:
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providing a blood sample;
extracting the hemoglobin from the sample and partially denaturing the hemoglobin;
exposing the hemoglobin to microparticles coated with zinc to allow binding of hemoglobin and glycated hemoglobin to the microparticles; and
determining the amount of glycated hemoglobin bound to the microparticles. - View Dependent Claims (17, 18, 19, 20, 21)
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22. An assay method for the in-vitro diagnosis and monitoring of diabetes mellitus through the assessment of glycated hemoglobin levels, the method comprising:
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providing a blood sample;
hemolysing the blood sample to release cell bounded hemoglobin and partially denature the hemoglobin;
capturing glycated hemoglobin on microparticles coated with zinc;
exposing the captured glycated hemoglobin on the microparticles to signal generating molecules which bind to the glycated hemoglobin;
removing signal generating molecules not bound to glycated hemoglobin; and
quantifying the amount of bound signal generating molecules to determine the percentage level of glycated hemoglobin present in the sample. - View Dependent Claims (23)
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24. A method of isolating hemoglobin from a biological sample, the method comprising:
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providing a biological sample;
capturing the hemoglobin on microparticles which are coated with zinc; and
eluting the microparticle-bound hemoglobin from the zinc coated microparticles. - View Dependent Claims (25, 26, 27)
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28. An assay for the selective binding of at least one target protein bearing a zinc binding site, comprising:
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providing a sample;
exposing the sample to zinc coated microparticles;
capturing the target proteins on the zinc coated microparticles;
isolating the microparticles;
detecting the presence of target proteins bound to the microparticles; and
optionally purifying the target protein from the microparticles. - View Dependent Claims (29)
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30. A zinc coated microparticle.
Specification