An ultrasensitive enzymatic method for measuring mevalonic acid in serum.
We have developed a simple, precise, and ultrasensitive enzymatic method for measuring serum mevalonic acid (MVA) concentration, which is thought to be a good indicator of the in vivo cholesterol biosynthesis rate. This assay is based on an enzyme cycling reaction and makes use of HMG-CoA reductase (HMGR), thio-NAD, NADH, and CoA. MVA participates in the HMGR cycling reaction, and its level is measured based on the production of thio-NADH, which is determined from the change in absorbance at 405 nm. To achieve high specificity, we used mevalonate kinase (MVK) in addition to HMGR. Only substrates able to participate in both the HMGR cycling reaction and the MVK reaction are measured as MVA. The detection limit for MVA is 0.4 ng/ml (2.7 nmol/l), and the calibration curve for MVA is linear up to 44 ng/ml (300 nmol/l). Regression analysis with 40 serum samples showed the accuracy of quantifying MVA with this enzymatic assay to be comparable to that using LC-MS/MS (correlation: y = 0.83x + 0.24; r = 0.97). This procedure is simple, precise, and robust. It is also rapid and has a high throughput, making it potentially useful for clinical applications.
Department of Diagnostics, Asahi Kasei Pharma Corporation, 632-1,Mifuku, Izunokuni-shi, Shizuoka 410-2321, Japan.
Takeshi
T
Matsuoka
Shigeru
S
Ueda
Hideyuki
H
Matsumoto
Masanobu
M
Kawakami
D016477:Artifacts; D001774:Blood Chemical Analysis / Q000379:methods*; D006801:Humans; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000097:blood; D057230:Limit of Detection; D016014:Linear Models; D008798:Mevalonic Acid / Q000097:blood* / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology; D015203:Reproducibility of Results; D013997:Time Factors
D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D006903:Hydroxymethylglutaryl CoA Reductases; D008798:Mevalonic Acid