Gorris Hans H, Soukka Tero
Department of Biochemistry, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic.
Department of Life Technologies/Biotechnology, University of Turku, Kiinamyllynkatu 10, 20520 Turku, Finland.
Anal Chem. 2022 Apr 26;94(16):6073-6083. doi: 10.1021/acs.analchem.1c05591. Epub 2022 Apr 11.
Immunoassays are important tools for clinical diagnosis as well as environmental and food analysis because they enable highly sensitive and quantitative measurements of analyte concentrations. In the 1980s, Roger Ekins suggested to improve the sensitivity of immunoassays by employing microspot assays, which are carried out under ambient analyte conditions and do not change the bulk analyte concentration of a sample during a measurement. More recently, the measurement of single analyte molecules has additionally attracted wide research interest. Although the ability to detect a single analyte molecule is not synonymous with the highest analytical sensitivity, single-molecule detection makes new routes accessible to avoiding background noise. This perspective follows the development of solid-phase immunoassays from the design of label techniques to single-molecule (digital) assays against the backdrop of Ekins's fundamental work on immunoassay theory. The essential aspects of both ambient analyte and digital assay approaches are presented as a guideline to finding a balance between the speed, sensitivity, and precision of immunoassays.
免疫测定是临床诊断以及环境和食品分析的重要工具，因为它们能够对分析物浓度进行高灵敏度和定量测量。20世纪80年代，罗杰·埃金斯建议通过采用微斑点测定法来提高免疫测定的灵敏度，这种测定法是在环境分析物条件下进行的，并且在测量过程中不会改变样品的总体分析物浓度。最近，对单个分析物分子的测量也引起了广泛的研究兴趣。虽然检测单个分析物分子的能力并不等同于最高的分析灵敏度，但单分子检测为避免背景噪声开辟了新途径。这一观点遵循了固相免疫测定从标记技术设计到单分子（数字）测定的发展历程，其背景是埃金斯在免疫测定理论方面的基础工作。文中介绍了环境分析物和数字测定方法的基本方面，作为在免疫测定的速度、灵敏度和精度之间找到平衡的指导方针。
