Proteins are involved in many biological processes. Misfolded, truncated, or mutated proteins as well as over- or underexpressed proteins have been implicated in many diseases. Therefore, detection and quantification of proteins is extremely important. Conventional techniques such as the enzyme-linked immunosorbent assay, Western Blot, and mass spectrometry have enabled discovery and study of proteins in biological samples. However, many important proteins are present at low concentrations, rendering them undetectable using conventional techniques. Furthermore, limited ability to simultaneously measure multiple proteins in a sample has constrained our ability to fully study the proteome. In this review, we comprehensively discuss approaches for protein detection. We first discuss the fundamentals of proteins and protein assays, including affinity reagents, surface functionalization, assay formats, signal detection, and multiplexing. We then discuss the challenges with these methods and review existing methods for highly sensitive and multiplexed protein detection. Finally, we review recent advances in protein detection from the literature and discuss challenges and future directions.

中文翻译：

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高灵敏度和多路复用的蛋白质测量

蛋白质参与许多生物学过程。错误折叠，截断或突变的蛋白质，以及过表达或表达不足的蛋白质都与许多疾病有关。因此，蛋白质的检测和定量非常重要。常规技术，例如酶联免疫吸附测定，蛋白质印迹法和质谱法，使得能够发现和研究生物样品中的蛋白质。但是，许多重要的蛋白质都以低浓度存在，这使得使用常规技术无法检测到它们。此外，同时测量样品中多种蛋白质的能力有限，这限制了我们全面研究蛋白质组的能力。在这篇综述中，我们全面讨论了蛋白质检测方法。我们首先讨论蛋白质和蛋白质测定的基础知识，包括亲和试剂，表面功能化，测定形式，信号检测和多路复用。然后，我们讨论了这些方法所面临的挑战，并回顾了用于高灵敏度和多重蛋白检测的现有方法。最后，我们从文献中回顾了蛋白质检测的最新进展，并讨论了挑战和未来方向。