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AssigneeBayer Aktiengesellschaft
InventorsDavid Sharpe, Anand Natrajan, Qingping Jiang, George Parsons, Say-Jong Law
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The present invention discloses a method for the measurement of hydride using a chemiluminescent compound. The preferred chemiluminescent molecule is an acridinium compound. The source of hydride for the reduction of acridinium compound may be of chemical or biochemical origin, or the result of enzymatic catalysis. The chemical source of hydride, for example, might be metal hydrides, such as NaBH4. A biochemical source of hydride might be that derived from NADH, or NADPH, while an enzymatic source would be the class of oxidoreductases termed dehydrogenases which convert NADH or NADPH from NAD or NADP. There are numerous potential applications for acridinium compounds as chemiluminescent indicators of hydride. Any applied tests or diagnostic assays, in which hydride is either present at the onset of or generated through the course of a reaction, would benefit from the present invention. Such tests, which could encompass many different formats as discussed below in detail, may involve the quantitation or detection of metal hydrides, or enzyme cofactors such as NADH, NADPH, FMNH2, or FADH2. Of particular importance, are those diagnostic assays which might use dehydrogenases as reagents, indicators, diagnostic markers or as labels. Ethanol, for example, might be detected with acridinium ester chemiluminescence through the reaction of alcohol dehydrogenase on ethanol, said reaction producing NADH. As a label, dehydrogenase might be used in an ELISA for the detection of a specific analyte with acridinium ester providing the signaling response. Nucleic acid assays using dehydrogenase as a label are also envisioned. Assays for the detection of clinically relevant dehydrogenases such as elevated glutamate dehydrogenase as an indicator of hepatocellular damage might also be developed.
A method for determining the amount of hydride produced in a reaction mixture wherein hydride is generated comprising (a) adding a known amount of a chemiluminescent compound to said reaction mixture, (b) adding reagents to said reaction mixture in step (a) wherein said reagents cause the release of chemiluminescence from said chemiluminescent compound, (c) measuring the amount of chemiluminescence given off from said reaction mixture after the addition of said reagents which release chemiluminescence, (d) comparing (1) the amount of chemiluminescence produced with (2) the expected chemiluminescence in the absence of said hydride to determine the difference in amount of chemiluminescence between (d)(1) and (d)(2), and (e) calculating the amount of said hydride present by comparison of said difference in chemiluminescence with a standard curve,
A method for determining the amount of hydride produced in a sample wherein hydride is generated, said method comprising (a) adding a known amount of a chemiluminescent compound to said sample, (b) adding reagents to said sample in step (a) which cause the release of chemiluminescence from said chemiluminescent compound, (c) measuring the amount of chemiluminescence given off from said sample after the addition of said reagents which release chemiluminescence, (d) comparing (1) the amount of chemiluminescence produced with (2) the expected chemiluminescence in the absence of said hydride to determine the difference in amount of chemiluminescence between (d)(1) and (d) (2), and (e) calculating the amount of said hydride present by comparison of said difference in chemiluminescence with a standard curve,
Dependencies are inferred from the claim text.