DETERMINATION OF GLYCATED PROTEIN
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Abstract
The present invention provides methods for quantitation of glycated protein in a biological sample using a solid support matrix by making a first bound protein measurement total bound protein under conditions where both glycated and non-glycated protein bind to the support in making a second bound protein measurement under conditions where glycated protein is bound to the support and non-glycated protein is not substantially bound. Diagnostic devices and kits comprising the methods of the present invention are also provided
72 Citations
60 Claims
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1-40. -40. (canceled)
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41. A method for determining a ratio of an amount of a glycated form of hemoglobin to a total amount of the hemoglobin in a blood sample, comprising:
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providing a solid support having negatively charged carboxyl groups immobilized thereon, which groups are capable of binding both the glycated and the nonglycated forms of the hemoglobin at a first pH, said support also having hydroxyboryl groups immobilized thereon, interspersed with the negatively charged carboxyl groups, which hydroxyboryl groups are capable of binding the glycated form of the hemoglobin at a second pH; lysing the sample; adding the sample to the solid support at the first pH, thereby binding both the glycated and the nonglycated forms of the hemoglobin to the negatively charged carboxyl groups on the solid support, and then performing a first measurement indicative of the total amount of the glycated and the nonglycated forms of the hemoglobin bound to the solid support; changing the pH on the support to the second pH, thereby removing both the nonglycated form of the hemoglobin and the glycated form of the hemoglobin from the negatively charged carboxyl groups, after which removal the glycated form of the hemoglobin binds to the hydroxyboryl groups on the solid support independent of incubation time, and then performing a second measurement indicative of the amount of the glycated form of the hemoglobin bound to the solid support; and determining the ratio of the amount of the glycated form of the hemoglobin to the total amount of the glycated and the nonglycated forms of the hemoglobin in the sample from the first and second measurements. - View Dependent Claims (42, 43, 44, 45, 46)
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47. A diagnostic device comprising:
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(1) a solid support matrix having a measurement area; and (2) negatively charged groups and dihydroxyboryl groups immobilized on the solid support matrix, wherein said negatively charged groups are capable of binding both glycated and nonglycated protein at a first pH, and said dihydroxyboryl groups are capable of binding glycated protein at a second pH. - View Dependent Claims (48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60)
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Specification