Methods for detecting lp-pla2 activity and inhibition of lp-pla2 activity
OwnerYaping Shou, Yin-Fai Siu +1inventors
InventorsYaping Shou, Yin-Fai Siu, George 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.
Claims23
A method for determining inhibition of Lp-PLA2 enzyme activity in at least one sample comprising the steps of preparing a solution comprising a substrate for Lp-PLA2 comprising a calorimetric or fluorometric detectable moiety;
- contacting at least one said sample with the solution of the preparing step;
- and
- detecting Lp-PLA2 activity, wherein the sample is from an animal that has been administered with Lp-PLA2 inhibitor.
A method for determining Lp-PLA2 enzyme activity in a sample obtained from an animal comprising the steps of:
- c) contacting 110 μL of a solution comprising:
- a solution comprising 90 mM 1-myristoryl-2-(4-nitrophenylsuccinyl) phosphatidylcholine contacted with a solution comprising 200 mM
- HEPES, 200 mM NaCl, 5 mM EDTA, 10 mM CHAPS, 10 mM sodium 1-nonanesulfonate at a pH 7.6 in a ratio of 0.66 μL to 110 μL;
- with at least one 25 μL tissue sample from an animal;
- with 25 μL each of a p-nitrophenol standard solution comprising;
- 4, 3, 2, 1, 0.4 or 0.2 nmol/μL p-nitrophenol in methanol;
- and
- 25 μL of phosphate buffered saline (PBS) or ddH2O to make a blank;
- and
- d) determining Lp-PLA2 activity.
Dependencies are inferred from the claim text.