Efficient Simulation of Arbitrary Multicomponent First-Order Binding Kinetics for Improved Assay Design and Molecular Assembly.
Efficient Simulation of Arbitrary Multicomponent First-Order Binding Kinetics for Improved Assay Design and Molecular Assembly.
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用于改进分析设计和分子组装的任意多组分一阶结合动力学的有效模拟。
DOI：
10.1021/acsmeasuresciau.1c00037
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发表时间：
2022-04-20
期刊：
影响因子：
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通讯作者：
Tabard-Cossa, Vincent
中科院分区：
文献类型：
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作者：
Briggs, Kyle;Bouhamidi, Mohamed Yassine;He, Liqun;Tabard-Cossa, Vincent
传统的酶联免疫吸附试验（ELISA）长期以来一直是利用微孔板检测特定靶蛋白的主力，但其灵敏度已接近基本极限。医疗保健领域的新机遇呼唤超高灵敏度的体外诊断试验。基于磁珠的三明治免疫分析格式已经开发出来，可以达到前所未有的灵敏度，比单个配体-受体相互作用的速率常数所允许的要好几个数量级。然而，这些超高灵敏度的分析容易受到许多混杂因素的影响，包括背景分子的非特异性结合和低丰度靶标在管壁和洗涤步骤中的丢失。此外，工作流的优化通常是耗时且昂贵的。在这项工作中，我们提出了一种模拟工具，允许用户以图形方式定义任意结合分析，包括完全可逆的一阶结合动力学，定时添加额外组分和定时洗涤步骤。该工具作为一个用户友好的web应用程序免费提供。该框架轻量级且快速，允许对任意复杂的分析方案进行廉价的模拟和可视化，包括但不限于数字免疫分析，DNA杂交和酶动力学，用于验证和优化分析设计，而不需要用户的任何编程知识。我们展示了其中的一些功能，并提供了分析模拟设计的实际指导。
Traditional enzyme-linked immunosorbent assay (ELISA), long the workhorse for specific target protein detection using microplate wells, is nearing its fundamental limit of sensitivity. New opportunities in health care call for in vitro diagnostic tests with ultrahigh sensitivity. Magnetic bead-based sandwich immunoassay formats have been developed that can reach unprecedented sensitivities, orders of magnitude better than are allowed for by the rate constants for a single ligand–receptor interaction. However, these ultrahigh sensitivity assays are vulnerable to a host of confounding factors, including nonspecific binding from background molecules and loss of low-abundance target to tube walls and during wash steps. Moreover, the optimization of workflow is often time-consuming and expensive. In this work, we present a simulation tool that allows users to graphically define arbitrary binding assays, including fully reversible first-order binding kinetics, timed addition of extra components, and timed wash steps. The tool is freely available as a user-friendly webapp. The framework is lightweight and fast, allowing for inexpensive simulation and visualization of arbitrarily complex assay schemes, including but not limited to digital immunoassays, DNA hybridization, and enzyme kinetics, for validation and optimization of assay designs without requiring any programming knowledge from the user. We demonstrate some of these capabilities and provide practical guidance on assay simulation design.