Pedersen Morten E, Østergaard Jesper, Jensen Henrik
FIDA-Tech Aps, C/O University of Copenhagen, Copenhagen, Denmark.
Department of Pharmacy, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2019;1972:109-123. doi: 10.1007/978-1-4939-9213-3_8.
Flow-Induced Dispersion Analysis (FIDA) enables characterization and quantification of proteins under native conditions. FIDA is based on measuring the change in size of a ligand as it selectively interacts with the target protein. The unbound ligand has a relatively small apparent hydrodynamic radius (size), which increase in the presence of the analyte due to binding to the analyte. The K of the interaction may be obtained in a titration experiment and the measurement of the apparent ligand size in an unknown sample forms the basis for determining the analyte concentration. The apparent molecular size is measured by Taylor dispersion analysis (TDA) in fused silica capillary capillaries. FIDA is a "ligand-binding" assay and has therefore certain features in common with Enzyme-Linked Immunosorbent Assay (ELISA), Surface Plasmon Resonance (SPR), and Biolayer Interferometry (BLI) based techniques. However, FIDA probes a single in-solution binding event and thus makes assay development straightforward, and the absolute size measurement enables built-in assay quality control. Further, as FIDA does not involve surface chemistries, complications related to nonspecific adsorption of analyte and assay components are minimized enabling direct measurement in, e.g., plasma and serum.
流动诱导分散分析（FIDA）能够在天然条件下对蛋白质进行表征和定量。FIDA基于测量配体与目标蛋白质选择性相互作用时其大小的变化。未结合的配体具有相对较小的表观流体动力学半径（大小），由于与分析物结合，在分析物存在时其大小会增加。相互作用的平衡解离常数（K）可在滴定实验中获得，未知样品中表观配体大小的测量构成了测定分析物浓度的基础。表观分子大小通过熔融石英毛细管中的泰勒分散分析（TDA）进行测量。FIDA是一种“配体结合”分析方法，因此与基于酶联免疫吸附测定（ELISA）、表面等离子体共振（SPR）和生物层干涉术（BLI）的技术有某些共同特征。然而，FIDA探测的是单个溶液内结合事件，因此使分析方法的开发变得简单直接，并且绝对大小测量实现了内置的分析质量控制。此外，由于FIDA不涉及表面化学，与分析物和分析组件非特异性吸附相关的复杂性被最小化，从而能够在例如血浆和血清中进行直接测量。
