Assay for glycated blood proteins
First Claim
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1. A method of assessing a glycated blood protein in a sample, said method comprising the steps of:
- a) optionally hemolyzing said sample to liberate cell bound glycated blood protein;
b) separating said glycated blood protein and the corresponding non-glycated blood protein from said sample using a liquid phase precipitation agent, or means substantially specific for the protein being assessed;
c) contacting said non-immobilized separated proteins with a first signal forming agent capable of binding to said glycated protein with substantially higher binding affinity than for the corresponding non-glycated protein;
d) optionally, contacting said non-immobilized separated proteins with a second signal forming agent capable of binding to said glycated protein and to said corresponding non-glycated protein; and
e) assessing the signal forming agents which have bound to said non-immobilized separated proteins or which have not bound to said glycated protein or said corresponding non-glycated protein;
with the proviso that where said glycated protein comprises glycated hemoglobin said first signal forming agent is a chromophore labelled boronic acid or salt thereof having an absorption maximum at above 600 nm.
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Abstract
A method of assessing glycated blood protein in a sample which comprises separating glycated and non-glycated protein using a liquid phase precipitation reagent, contacting the sample with a signal forming agent capable of binding preferentially to the glycated protein, and assessing the signal forming agents.
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Citations
28 Claims
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1. A method of assessing a glycated blood protein in a sample, said method comprising the steps of:
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a) optionally hemolyzing said sample to liberate cell bound glycated blood protein; b) separating said glycated blood protein and the corresponding non-glycated blood protein from said sample using a liquid phase precipitation agent, or means substantially specific for the protein being assessed; c) contacting said non-immobilized separated proteins with a first signal forming agent capable of binding to said glycated protein with substantially higher binding affinity than for the corresponding non-glycated protein; d) optionally, contacting said non-immobilized separated proteins with a second signal forming agent capable of binding to said glycated protein and to said corresponding non-glycated protein; and e) assessing the signal forming agents which have bound to said non-immobilized separated proteins or which have not bound to said glycated protein or said corresponding non-glycated protein; with the proviso that where said glycated protein comprises glycated hemoglobin said first signal forming agent is a chromophore labelled boronic acid or salt thereof having an absorption maximum at above 600 nm. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14)
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15. A method of assessing a glycated blood protein in a sample, said method comprising the steps of:
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a) optionally hemolyzing said sample to liberate cell bound glycated blood protein; b) separating said glycated blood protein and the corresponding non-glycated blood protein from said sample using a liquid phase precipitation agent, or means substantially specific for the protein being assessed; c) subsequently contacting said non-immobilized separated proteins with a first signal forming agent capable of binding to said glycated protein with substantially higher binding affinity than for the corresponding non-glycated protein; d) optionally, contacting said non-immobilized separated proteins with a second signal forming agent capable of binding to said glycated protein and to said corresponding non-glycated protein; and e) assessing the signal forming agents which have bound to said non-immobilized separated proteins and which have not bound to said glycated protein or said corresponding non-glycated protein; with the proviso that where said glycated protein comprises glycated hemoglobin said first signal forming agent is a chromophore labelled boronic acid or salt thereof having an absorption maximum at above 600 nm. - View Dependent Claims (16, 17, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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- 18. A method as claimed in claim 18 wherein precipitation is achieved by addition of metallic cations.
Specification