Heart Institute, Cedars-Sinai Medical Center , 127 South San Vicente Boulevard, Los Angeles, California 90048, United States.
Sciex Separations , 250 South Kraemer Boulevard, Brea, California 92821, United States.
J Proteome Res. 2017 Dec 1;16(12):4515-4522. doi: 10.1021/acs.jproteome.7b00482. Epub 2017 Oct 2.
B-type Natriuretic Peptide (BNP) is a biologically active circulating hormone. Plasma concentrations of BNP are routinely used in the diagnosis of heart failure, and the intravenous infusion of recombinant BNP can be used for heart failure treatment. Like many bioactive polypeptides, multiple plasma enzymes are known to cleave circulating BNP, and as part of the CVD-B/D-HPP mandate, we sought to develop a technique capable of profiling these catabolic processes in plasma. We used a neutral-coated capillary electrophoresis-electrospray ionization (CESI) separation system coupled with high-resolution mass spectrometry to profile the proteolysis of exogenous recombinant BNP in plasma. Our method utilizes electrokinetic injection of minimally processed plasma samples to simultaneously monitor the dynamic generation and breakdown of at least five BNP peptidoforms in plasma. By integrating multisegment injection, our method can produce a multipoint BNP proteolytic profile for one sample within an hour. We envision applying this method to assess the potential relation between plasma-based BNP proteolysis and heart failure as well as a means of monitoring BNP bioavailability after therapeutic infusion.
B 型利钠肽（BNP）是一种生物活性循环激素。血浆 BNP 浓度通常用于心力衰竭的诊断，重组 BNP 的静脉输注可用于心力衰竭的治疗。与许多生物活性多肽一样，已知多种血浆酶可切割循环 BNP，作为 CVD-B/D-HPP 任务的一部分，我们试图开发一种能够对这些血浆代谢过程进行分析的技术。我们使用中性涂层毛细管电泳-电喷雾电离（CESI）分离系统结合高分辨率质谱法来分析血浆中外源重组 BNP 的蛋白水解。我们的方法利用电动进样最小处理的血浆样品，同时监测至少五种 BNP 肽形式在血浆中的动态生成和分解。通过整合多段进样，我们的方法可以在一个小时内为一个样本生成多点 BNP 蛋白水解谱。我们设想应用这种方法来评估基于血浆的 BNP 蛋白水解与心力衰竭之间的潜在关系，以及监测治疗性输注后 BNP 生物利用度的一种手段。
