Sucrose detection by enzyme-linked immunosorbant assay

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 14, 435 15, 435 18, 435178, 435210, 435 732, 435 71, 435 704, 436518, 436524, G01N 33533

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active

059726318

ABSTRACT:
A method is described for the rapid, sensitive and accurate determination of sucrose in biological fluids. A substrate is pre-coated with a glucose or fructose polymer and a transglycosidase enzyme. When the coated substrate is incubated with biological fluids containing concentrations of sucrose, the transglycosidase enzyme transfers monomers of glucose or fructose from the sucrose to the glucose or fructose polymer. The dimensions of the polymer are increased in proportion to the sucrose concentration of the samples. Newly formed polymer is subsequently quantitated in an immunoassay which employs either a combination of a carbohydrate-binding protein (which may be an antibody) and a conjugate of a secondary antibody and a marker enzyme, or a conjugate of a carbohydrate-binding protein and a marker enzyme. The assay is accurate at sucrose concentrations below 1 .mu.g/mL. No interference was observed at glucose concentrations as high as 25 mM. The sucrose detection assay is particularly useful in a non-invasive diagnostic test for gastric damage.

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