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Tutorial: design and fabrication of nanoparticle-based lateral-flow immunoassays
Nature Protocols ( IF 14.8 ) Pub Date : 2020-10-23 , DOI: 10.1038/s41596-020-0357-x
Claudio Parolo 1 , Amadeo Sena-Torralba 1 , José Francisco Bergua 1 , Enric Calucho 1 , Celia Fuentes-Chust 1 , Liming Hu 1 , Lourdes Rivas 1 , Ruslan Álvarez-Diduk 1 , Emily P Nguyen 1 , Stefano Cinti 2 , Daniel Quesada-González 3 , Arben Merkoçi 1, 4
Affiliation  

Lateral-flow assays (LFAs) are quick, simple and cheap assays to analyze various samples at the point of care or in the field, making them one of the most widespread biosensors currently available. They have been successfully employed for the detection of a myriad of different targets (ranging from atoms up to whole cells) in all type of samples (including water, blood, foodstuff and environmental samples). Their operation relies on the capillary flow of the sample throughout a series of sequential pads, each with different functionalities aiming to generate a signal to indicate the absence/presence (and, in some cases, the concentration) of the analyte of interest. To have a user-friendly operation, their development requires the optimization of multiple, interconnected parameters that may overwhelm new developers. In this tutorial, we provide the readers with: (i) the basic knowledge to understand the principles governing an LFA and to take informed decisions during lateral flow strip design and fabrication, (ii) a roadmap for optimal LFA development independent of the specific application, (iii) a step-by-step example procedure for the assembly and operation of an LF strip for the detection of human IgG and (iv) an extensive troubleshooting section addressing the most frequent issues in designing, assembling and using LFAs. By changing only the receptors, the provided example procedure can easily be adapted for cost-efficient detection of a broad variety of targets.



中文翻译:

教程:基于纳米粒子的横向流动免疫分析的设计和制造

侧向流动分析 (LFA) 是一种快速、简单且廉价的分析方法,可在护理点或现场分析各种样品,使其成为目前可用的最广泛的生物传感器之一。它们已成功用于检测所有类型样品(包括水、血液、食品和环境样品)中的无数不同目标(从原子到整个细胞)。它们的操作依赖于样品在一系列连续垫中的毛细流动,每个垫都具有不同的功能,旨在生成信号以指示感兴趣的分析物的不存在/存在(在某些情况下,还包括浓度)。为了实现用户友好的操作,他们的开发需要优化多个相互关联的参数,这可能会让新开发人员不知所措。在本教程中,我们为读者提供:(i) 了解 LFA 管理原则的基本知识,并在横向流动带设计和制造过程中做出明智的决定,(ii) 独立于特定应用的最佳 LFA 开发路线图,(iii)用于检测人类 IgG 的 LF 条的组装和操作的分步示例程序,以及 (iv) 解决设计、组装和使用 LFA 中最常见问题的广泛故障排除部分。通过仅更改受体,所提供的示例程序可以很容易地适用于对各种目标进行经济高效的检测。(ii) 独立于特定应用的最佳 LFA 开发路线图,(iii) 用于检测人 IgG 的 LF 条带的组装和操作的分步示例程序,以及 (iv) 广泛的故障排除部分解决设计、组装和使用 LFA 时最常见的问题。通过仅更改受体,所提供的示例程序可以很容易地适用于对各种目标进行经济高效的检测。(ii) 独立于特定应用的最佳 LFA 开发路线图,(iii) 用于检测人 IgG 的 LF 条带的组装和操作的分步示例程序,以及 (iv) 广泛的故障排除部分解决设计、组装和使用 LFA 时最常见的问题。通过仅更改受体,所提供的示例程序可以很容易地适用于对各种目标进行经济高效的检测。

更新日期:2020-10-28
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