Determination of glycoprotein and glycosylated hemoglobin in blo

Electrolysis: processes – compositions used therein – and methods – Electrolytic analysis or testing – Using ion exchange resin

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205778, 204403, G01N 2726

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active

060540397

ABSTRACT:
A method of determining the concentration of glycoproteins and glycosylated hemoglobin in whole blood and whole blood components by means of an amperometric biosensor and an amperometric biosensor for this determination are provided. In one embodiment, whole blood is introduced into a version of an amperometric sensor having a component that removes erythrocytes. Redox mediators are used to obtain a current flow based on the oxidation of fructosamine derivatives that can be correlated with the concentration of glycosylated proteins in the fraction of the blood from which erythrocytes have been excluded. To obtain the concentration of glycosylated hemoglobin, whole blood is introduced into a version of the sensor which includes a component that produces lysis of the erythrocytes yielding a current flow proportional to the total quality of glycosylated proteins including glycosylated hemoglobin. The glycosyltaed hemoglobin concentration is obtained by subtracting the glycoprotein concentration in the absence of erythrocytes from the glycoprotein concentration of the lysed whole blood. The sensor generally comprises a sensing electrode having a first redox mediator dispersed in an electrically conductive medium such as an electrically conductive graphite formulation; a reference electrode such as a standard silver-silver chloride electrode; a reagent strip containing a pH buffer and a second redox mediator system in a gel medium; and a whole blood treatment component consisting of either a membrane or other means to filter erythrocytes from whole blood or a means to lyse erythrocytes. In a preferred form, that has high sensitivity, the sensing electrode and the reference electrode may be formed as coatings on separate non-conductive strips such as polyester film with these strips arranged so that they form "the bread" of a sandwich in which the electrode coatings are face-to-face and the reagent strip and the filtration or lysing component form the "filling" of the sandwich. The filtration or cell lysing component covers an opening in the reference electrode through which samples are introduced, and is superimposed on the reagent strip.

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Johnson, et al, "Fructosamine: a new approach to the estimation of serum glycosylprotein. An index of diabetic control" Clinica Chimica Acta, 127 (1982) 87-95 month unknown.

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