Quantification of NTproBNP in Rat Serum Using Immunoprecipitation and LC/MS/MS: a Biomarker of Drug-Induced Cardiac Hypertrophy

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• 作者：Michael Berna ; Lee Ott ; Steve Engle ; David Watson ; Philip Solter ; Bradley Ackermann
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：February 1, 2008
• 年：2008
• 卷：80
• 期：3
• 页码：561 - 566
• 全文大小：197K
• 年卷期：v.80,no.3(February 1, 2008)
• ISSN：1520-6882

文摘

Brain natriuretic peptide (BNP) and N-terminal proBNP(NTproBNP) are well established in the clinic as biomarkers of heart failure. BNP hormone and the inactiveNTproBNP are predominantly secreted in the ventriclesof the heart in response to pressure overload and,consequently, are being investigated as markers of drug-induced cardiac hypertrophy in rat to support drugdevelopment. In the work presented here, an immunoaffinity-based LC/MS/MS assay was developed and validated to measure a selective tryptic fragment of NTproBNP in rat serum. The assay covers the range of 13-329pg/mL of the tryptic fragment LLELIR, corresponding to0.1-2.5 ng/mL intact NTproBNP. A stable isotope-labeled version of NTproBNP containing the tryptic fragment LLELI[¹³C₆¹⁵N₁]R was prepared by solid-phasepeptide synthesis and was used as an internal standardto minimize assay variability. Due to endogenous NTproBNP present in rat serum, human serum was usedas the control matrix, and parallelism between rat andhuman serum was established by standard addition.Assay accuracy (% RE) and precision (% CV) weremeasured at three concentrations on each of 4 days anddid not exceed 4.2 and 14.5%, respectively. Additionally,study data are presented from the application of this assayin which rats demonstrated a significant increase inNTproBNP serum concentration following administrationof an agent known to induce cardiac hypertrophy. In thisstudy, the relationship between serum NTproBNP andcardiac hypertrophy was corroborated by increases inheart weight and magnetic resonance imaging of the testsubjects' left ventricle. To our knowledge, this representsthe first reported assay for NTproBNP in preclinicalspecies for the assessment of cardiac hypertrophy.