Measurement of glycosylated hemoglobin by immunoassay

Details Measurement of glycosylated hemoglobin by immunoassay Measurement of glycosylated hemoglobin by immunoassay

1987-02-05

1989-02-21

Kepplinger, Esther M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

435 28, 435 68, 435188, 435810, 436 67, 436175, 436811, G01N 5300, G01N 3372, G01N 100, C12Q 128

Patent

active

048064688

ABSTRACT:
A method for determining glycosylated hemoglobin in a diluted blood sample includes binding of glycosylated hemoglobin to a specific monoclonal antibody and binding of nonglycosylated hemoglobin to a polyclonal antibody. The polyclonal antibody binds to all hemoglobin fractions and blocks the peroxidase activity thereof, except glycosylated hemoglobin bound to the monoclonal antibody which retains peroxidase activity. A beam of light having a wavelength of about 416 nm is passed through the assay medium and is absorbed by all hemoglobin fractions, bound or unbound, and this absorbance thereby provides a measure of total hemoglobin. Hydrogen peroxide and a peroxidase substrate are added. The substrate is oxidized by the peroxide to a product having an absorbance at a wavelength different from 416 nm, the oxidation being catalyzed by the retained peroxidase activity of the bound glycosylated hemoglobin. This absorbance provides a measure of glycosylated hemoglobin. From the two absorbances, the percentage of glycosylated hemoglobin in the total hemoglobin of the sample may be calculated. The invention includes a kit of materials useful in performing the assay method of the invention.

REFERENCES:
patent: 4582811 (1986-04-01), Pucci et al.
patent: 4695552 (1987-09-01), Schmitt et al.
patent: 4727036 (1988-02-01), Knowles et al.

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