Methods for detecting LP-PLA2 activity and inhibition of LP-PLA2 activity
AssigneeGlaxo Group Limited
InventorsYaping Shou, Yin-Fai Siu, George T. Walker
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Abstract
This invention relates to methods for determining the activity of Lp-PLA2 in at least one sample from an animal. The invention also relates to methods for determining the inhibition of Lp-PLA2 activity in samples from animals that are administered an Lp-PLA2 inhibitor.
Claims29
An Lp-PLA2 colorimetric activity assay mixture that can detect Lp-PLA2 activity in the presence of an inhibitor of Lp-PLA2 activity, the assay mixture made by combining:
- a substrate solution comprising 1-myristoyl-2-(p-nitrophenylsuccinyl)phosphatidylcholine;
- a buffer solution comprising a detergent;
- and
- a plasma sample comprising an inhibitor of Lp-PLA2, wherein the plasma sample has been diluted about 3 to 9 fold in the assay mixture, and
- wherein the concentration of substrate in the assay mixture is less than or equal to about the Km of the substrate.
An Lp-PLA2 colorimetric activity assay mixture that can detect Lp-PLA2 activity in the presence of an inhibitor of Lp-PLA2 activity, the assay mixture made by combining:
- a substrate solution comprising 1-myristoyl-2-(p-nitrophenylsuccinyl)phosphatidylcholine,
- a buffer solution comprising a detergent, and
- a plasma sample comprising an inhibitor of Lp-PLA2, wherein the plasma sample has been diluted about 3 to 9 fold in the assay mixture, and
- wherein the concentration of substrate is between about 53 μM and about 1125 μM in the assay mixture.
A colorimetric method of determining inhibition of Lp-PLA2 enzyme activity in a sample of blood plasma obtained from a human patient taking an inhibitor of Lp-PLA2, the method comprising:
- forming an assay mixture by combining a plasma sample with a buffer solution comprising a detergent and a substrate solution comprising 1-myristoyl-2-(p-nitrophenylsuccinyl)phosphatidylcholine so that the plasma sample is diluted about 3 to about 9 fold and the concentration of substrate in the assay mixture is less than or equal to the Km of the substrate;
- and
- colorimetrically sampling the assay mixture to determine activity of the Lp-PLA2 enzyme.
A colorimetric method of determining inhibition of Lp-PLA2 enzyme activity in a sample of blood plasma obtained from a human patient taking an inhibitor of Lp-PLA2, the method comprising:
- forming an assay mixture by combining a plasma sample from a patient with a buffer solution comprising a detergent and a substrate solution comprising 1-myristoyl-2-(p-nitrophenylsuccinyl)phosphatidylcholine so that the plasma sample is diluted about 3 to about 9 fold and the concentration of substrate is between about 53 μM to about 1125 μM in the assay mixture;
- and
- colorimetrically sampling to determine an absorbance from the colorimetric sample solution.
Dependencies are inferred from the claim text.