Mass spectrometry assay for plasma-renin
AssigneeQuest Diagnostics Investments Incorporated
InventorsCory E. Bystrom, Richard E. Reitz, Nigel J. Clarke
Life of this patent
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Abstract
Provided are methods for measuring renin activity in a plasma sample using mass spectrometry. The methods generally involve ionizing purified angiotensin 1 from the sample and detecting the amount of angiotensin 1 ions generated. The amount of detected angiotensin 1 ions are then related to the amount of angiotensin 1 generated in the sample, which in turn is related to renin activity in the sample.
Claims43
A method for measuring the amount of angiotensin 1 in a sample, said method comprising:
- (a) purifying angiotensin 1 from said sample by solid-phase extraction;
- (b) ionizing angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (c) detecting the amount of the angiotensin ion(s) by mass spectrometry, said ions selected from the group consisting of ions with a mass/charge ratio of 433.0±0.5, 619.5±0.5, 647.4±0.5 and 1297±0.5;
- and
- (d) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample.
A method for measuring the amount of angiotensin 1 in a sample, said method comprising:
- (a) incubating said sample premixed with aminoethylbenzylsulfonyl fluoride under conditions suitable for the generation of angiotensin 1 by renin in the sample;
- (b) purifying angiotensin 1 in said sample by liquid chromatography;
- (c) ionizing the purified angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (d) detecting the amount of the angiotensin 1 ion(s) by mass spectrometry;
- and
- (e) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample.
A method for measuring the amount of angiotensin 1 in a sample, said method comprising:
- (a) incubating the sample premixed with a water stable protease inhibitor that is not effective against renin under conditions suitable for the generation of angiotensin 1 by renin in the sample;
- (b) purifying angiotensin 1 in said sample by solid phase extraction;
- (c) further purifying angiotensin 1 in said sample by liquid chromatography with on-line processing;
- (d) ionizing the purified angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (e) detecting the amount of the angiotensin 1 ion(s) by mass spectrometry,
- (f) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample.
A method for measuring renin activity in a sample, said method comprising:
- (a) incubating the sample premixed with a water stable protease inhibitor that is not effective against renin under conditions suitable for the generation of angiotensin 1 by renin in the sample;
- (b) purifying angiotensin 1 from said sample by solid phase extraction;
- (c) further purifying angiotensin 1 by liquid chromatography with on-line processing;
- (d) ionizing the purified angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (e) detecting the amount of the angiotensin 1 ion(s) by mass spectrometry,
- (f) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample;
- and
- (g) using the amount of angiotensin 1 measured in said sample to calculate renin activity in the sample.
A method for measuring renin activity in a sample, said method comprising,
- (a) incubating the sample premixed with aminoethylbenzylsulfonyl fluoride under conditions suitable for the generation of angiotensin 1 by renin in the sample;
- (b) purifying angiotensin 1 by liquid chromatography;
- (c) ionizing the purified angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (d) detecting the amount of the angiotensin 1 ion(s) by mass spectrometry;
- (e) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample;
- and
- (f) using the amount of angiotensin 1 measured in said sample to calculate renin activity in the sample.
A method for measuring renin activity in a sample, said method comprising,
- (a) incubating the sample premixed with a water stable protease inhibitor that is not effective against renin under conditions suitable for the generation of angiotensin 1 by renin in the sample;
- (b) purifying angiotensin 1 in said sample by liquid chromatography;
- (c) ionizing the purified angiotensin 1 from said sample to produce one or more ions detectable by mass spectrometry;
- (d) detecting the amount of the angiotensin 1 ion(s) by mass spectrometry, said ions selected from the group consisting of ions with a mass/charge ratio of 433.0±0.5, 619.5±0.5, 647.4±0.5 and 1297±0.5;
- and
- (e) using the amount of ion(s) detected to measure the amount of angiotensin 1 in said sample:
- and
- (f) using the amount of angiotensin 1 measured in said sample to calculate renin activity in the sample.
Dependencies are inferred from the claim text.