Bioanalytical Mass Spectrometry Facility, University of New South Wales, Sydney, NSW, Australia.
Centre for Healthy Brain Ageing, School of Psychiatry, University of New South Wales, Sydney, NSW, Australia.
Methods Mol Biol. 2020;2138:49-81. doi: 10.1007/978-1-0716-0471-7_3.
The apolipoproteins are well known for their roles in both health and disease, as components of plasma lipoprotein particles, such as high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), chylomicrons, and metabolic, vascular- and inflammation-related disorders, such as cardiovascular disease, atherosclerosis, metabolic syndrome, and diabetes. Increasingly, their roles in neurovascular and neurodegenerative disorders are also being elucidated. They play major roles in lipid and cholesterol transport between blood and organs and are, therefore, critical to maintenance and homeostasis of the lipidome, with apolipoprotein-lipid interactions, including cholesterol, fatty acids, triglycerides, phospholipids, and isoprostanes. Further, they have important pleiotropic roles related to aging and longevity, which are largely managed through their many structural variants, including multiple isoforms, and a diversity of post-translational modifications. Consequently, tools for the characterization and accurate quantification of apolipoproteins, including their diverse array of variant forms, are required to understand their salutary and disease related roles. In this chapter we outline three distinct quantitative approaches suitable for targeting apolipoproteins: (1) multiplex immunoassays, (2) mass spectrometric immunoassay, and (3) multiple reaction monitoring, mass spectrometric quantification. We also discuss management of pre-analytical and experimental design variables.
载脂蛋白在健康和疾病中都起着重要的作用，它们是血浆脂蛋白颗粒（如高密度脂蛋白（HDL）、低密度脂蛋白（LDL）、极低密度脂蛋白（VLDL）、乳糜微粒）的组成部分，与代谢、血管和炎症相关的疾病（如心血管疾病、动脉粥样硬化、代谢综合征和糖尿病）有关。越来越多的证据表明，它们在神经血管和神经退行性疾病中的作用也正在被阐明。它们在血液和器官之间的脂质和胆固醇转运中起着重要作用，因此对脂质组的维持和平衡至关重要，载脂蛋白-脂质相互作用包括胆固醇、脂肪酸、甘油三酯、磷脂和异前列烷。此外，它们在衰老和长寿方面具有重要的多效性作用，这些作用主要通过它们的许多结构变体来管理，包括多种同工型和多种翻译后修饰。因此，需要用于鉴定和准确定量载脂蛋白的工具，包括它们多样化的变体形式，以了解其有益和与疾病相关的作用。在本章中，我们概述了三种不同的定量方法，适用于靶向载脂蛋白：（1）多重免疫分析，（2）质谱免疫分析，（3）多重反应监测，质谱定量。我们还讨论了分析前和实验设计变量的管理。
