Baldelli Elisa, Calvert Valerie, Hodge Alex, VanMeter Amy, Petricoin Emanuel F, Pierobon Mariaelena
Center for Applied Proteomics and Molecular Medicine, George Mason University, 10920 George Mason Circle, MS 1A9, Manassas, VA, 20110, USA.
Methods Mol Biol. 2017;1606:149-169. doi: 10.1007/978-1-4939-6990-6_11.
While genes and RNA encode information about cellular status, proteins are considered the engine of the cellular machine, as they are the effective elements that drive all cellular functions including proliferation, migration, differentiation, and apoptosis. Consequently, investigations of the cellular protein network are considered a fundamental tool for understanding cellular functions.Alteration of the cellular homeostasis driven by elaborate intra- and extracellular interactions has become one of the most studied fields in the era of personalized medicine and targeted therapy. Increasing interest has been focused on developing and improving proteomic technologies that are suitable for analysis of clinical samples. In this context, reverse-phase protein microarrays (RPPA) is a sensitive, quantitative, high-throughput immunoassay for protein analyses of tissue samples, cells, and body fluids.RPPA is well suited for broad proteomic profiling and is capable of capturing protein activation as well as biochemical reactions such as phosphorylation, glycosylation, ubiquitination, protein cleavage, and conformational alterations across hundreds of samples using a limited amount of biological material. For these reasons, RPPA represents a valid tool for protein analyses and generates data that help elucidate the functional signaling architecture through protein-protein interaction and protein activation mapping for the identification of critical nodes for individualized or combinatorial targeted therapy.
虽然基因和RNA编码有关细胞状态的信息，但蛋白质被认为是细胞机器的引擎，因为它们是驱动包括增殖、迁移、分化和凋亡在内的所有细胞功能的有效元件。因此，细胞蛋白质网络的研究被认为是理解细胞功能的基本工具。由复杂的细胞内和细胞外相互作用驱动的细胞稳态改变已成为个性化医学和靶向治疗时代研究最多的领域之一。人们越来越关注开发和改进适用于临床样本分析的蛋白质组学技术。在这种背景下，反相蛋白质微阵列（RPPA）是一种用于组织样本、细胞和体液蛋白质分析的灵敏、定量、高通量免疫测定法。RPPA非常适合广泛的蛋白质组学分析，并且能够使用有限量的生物材料捕获数百个样本中的蛋白质激活以及生化反应，如磷酸化、糖基化、泛素化、蛋白质裂解和构象改变。由于这些原因，RPPA是蛋白质分析的有效工具，并生成有助于通过蛋白质-蛋白质相互作用和蛋白质激活图谱阐明功能信号架构的数据，以识别个性化或联合靶向治疗的关键节点。
