Measurement of plasma renin concentration in cats by use of a fluorescence resonance energy transfer peptide substrate of renin

Chad W. Schmiedt Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

Search for other papers by Chad W. Schmiedt in
Current site
Google Scholar
PubMed
Close
 DVM
,
Katherine A. E. Hurley Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

Search for other papers by Katherine A. E. Hurley in
Current site
Google Scholar
PubMed
Close
,
Xiaohe Tong AnaSpec Inc, 34801 Campus Dr, Fremont, CA 94555.

Search for other papers by Xiaohe Tong in
Current site
Google Scholar
PubMed
Close
 MS
,
Vera A. Rakhmanova AnaSpec Inc, 34801 Campus Dr, Fremont, CA 94555.

Search for other papers by Vera A. Rakhmanova in
Current site
Google Scholar
PubMed
Close
 PhD
,
Cecilia L. Po AnaSpec Inc, 34801 Campus Dr, Fremont, CA 94555.

Search for other papers by Cecilia L. Po in
Current site
Google Scholar
PubMed
Close
 BS
, and
David J. Hurley Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.
Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602.

Search for other papers by David J. Hurley in
Current site
Google Scholar
PubMed
Close
 PhD
Restricted access
Purchase Article

Abstract

Objective—To evaluate the use of a commercially available 5-carboxyfluorescein–based, intramolecularly quenched, fluorescence resonance energy transfer (FRET) peptide substrate of renin for measurement of plasma renin concentration in cats.

Sample Population—Plasma samples obtained during a previous study of renal autograft ischemia-reperfusion injury in 10 cats and samples of fetal bovine serum containing recombinant human renin (rh-renin).

Procedures—Experiments involving samples of fetal bovine serum containing rh-renin were conducted to identify a suitable control vehicle, optimal substrate concentration, and appropriate duration of incubation. With the use of the identified assay conditions, a standard curve was constructed to allow conversion of relative fluorescent units into values of renin concentration (ng/mL). Subsequently, plasma samples obtained from cats before and after renal autograft ischemia-reperfusion injury were assayed to determine endogenous renin concentration.

Results—Under conditions of a 1:50 substrate dilution and 4-hour incubation period, the assay detected small amounts of rh-renin in fetal bovine serum. A linear relationship (R2 = 0.996) between the relative fluorescent units generated and exogenous rh-renin concentration was evident. The assay detected renin in plasma samples obtained from cats after renal autograft ischemia-reperfusion, and renin concentrations on days 1 and 2 after transplant differed significantly.

Conclusions and Clinical Relevance—The study data indicated that the assay involving the FRET peptide substrate of renin is potentially a rapid and specific method for measurement of plasma renin concentration in cats.

All Time Past Year Past 30 Days
Abstract Views 34 0 0
Full Text Views 1825 1246 150
PDF Downloads 245 92 6
Advertisement