ICON Bioanalytical Laboratories, Amerikaweg 18, 9407 TK Assen, the Netherlands; Department of Analytical Biochemistry University of Groningen, A. Deusinglaan 1, 9700 AV Groningen, the Netherlands; Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, EA61, P.O. Box 30.001, 9700 RB Groningen, the Netherlands.
Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, EA61, P.O. Box 30.001, 9700 RB Groningen, the Netherlands.
Clin Chim Acta. 2024 Feb 1;554:117736. doi: 10.1016/j.cca.2023.117736. Epub 2023 Dec 22.
An LC-MS/MS method is presented for the simultaneous quantification of two structurally closely related protein biomarker isoforms, the 22-kDa isoforms of human growth hormone 1 and human growth hormone 2, in human plasma. It is based on multiplexed immunocapture using two monoclonal antibodies immobilized on magnetic beads, tryptic digestion and quantification of two specific signature peptides plus an additional peptide for estimation of total growth hormone related concentrations. A full validation according to international guidelines was performed across the clinically relevant concentration ranges of 0.5 to 50 ng/mL for growth hormone 1, and 2 to 50 ng/mL for growth hormone 2 and demonstrated satisfactory method performance in terms of accuracy, precision, stability and absence of interference. The method's applicability for routine analysis and its ability to effectively distinguish between GH1 and GH2 was demonstrated by the analysis of plasma samples from pregnant individuals to study the changes in growth hormone levels during pregnancy.
一种 LC-MS/MS 方法被提出，用于同时定量两种结构上密切相关的蛋白质生物标志物同工型，即人类生长激素 1 和人类生长激素 2 的 22kDa 同工型，在人血浆中。它基于使用固定在磁性珠上的两种单克隆抗体的多重免疫捕获，胰蛋白酶消化和两种特定特征肽的定量，外加一个用于估计总生长激素相关浓度的肽。根据国际指南进行了全面验证，涵盖了生长激素 1 的临床相关浓度范围为 0.5 至 50ng/mL，生长激素 2 的浓度范围为 2 至 50ng/mL，在准确性、精密度、稳定性和无干扰方面表现出令人满意的方法性能。该方法通过分析来自孕妇的血浆样本，以研究怀孕期间生长激素水平的变化，证明了其在常规分析中的适用性及其有效区分 GH1 和 GH2 的能力。
