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用于多重消费免疫诊断的可互连固液蛋白质提取单元及基于芯片的稀释技术。

Interconnectable solid-liquid protein extraction unit and chip-based dilution for multiplexed consumer immunodiagnostics.

作者信息

Ross Georgina M S, Filippini Daniel, Nielen Michel W F, Salentijn Gert Ij

机构信息

Wageningen Food Safety Research (WFSR), Wageningen University & Research, P.O. Box 230, 6700, AE, Wageningen, the Netherlands.

Optical Devices Laboratory, Division of Sensor and Actuator Systems, IFM - Linköping University, S58183, Linköping, Sweden.

出版信息

Anal Chim Acta. 2020 Dec 15;1140:190-198. doi: 10.1016/j.aca.2020.10.018. Epub 2020 Oct 16.

Abstract

While consumer-focused food analysis is upcoming, the need for multiple sample preparation and handling steps is limiting. On-site and consumer-friendly analysis paradoxically still requires laboratory-based and skill-intensive sample preparation methods. Here, we present a compact, inexpensive, and novel prototype immunosensor combining sample preparation and on-chip reagent storage for multiplex allergen lateral flow immunosensing. Our comprehensive approach paves the way for personalized consumer diagnostics. The prototype allows for handheld solid-liquid extraction, pipette-free on-chip dilution, and adjustment of sample concentrations into the appropriate assay dynamic working range. The disposable and interconnectable homogenizer unit allows for the extraction and 3D-sieve based filtration of allergenic proteins from solid bakery products in 1 min. The homogenizer interconnects with a 3D-printed unibody lab-on-a-chip (ULOC) microdevice, which is used to deliver precise volumes of sample extract to a reagent reservoir. The reagent reservoir is implemented for on-chip storage of carbon nanoparticle labeled antibodies and running buffer for dilution. The handheld prototype allows for total homogenization of solid samples, solid-liquid protein extraction, 3D-printed sieve based filtration, ULOC-enabled dilution, mixing, transport, and smartphone-based detection of hazelnut and peanut allergens in solid bakery products with limited operational complexity. The multiplex lateral flow immunoassay (LFIA) detects allergens as low as 0.1 ppm in real bakery products, and the system is already consumer-operable, demonstrating its potential for future citizen science approaches. The designed system is suitable for a wide range of analytical applications outside of food safety, provided an LFIA is available.

摘要

虽然以消费者为中心的食品分析即将出现,但多个样品制备和处理步骤的需求限制了其发展。矛盾的是,现场和消费者友好型分析仍然需要基于实验室且技术密集型的样品制备方法。在此,我们展示了一种紧凑、廉价且新颖的免疫传感器原型,它结合了样品制备和片上试剂存储功能,用于多重过敏原侧向流动免疫传感。我们的综合方法为个性化消费者诊断铺平了道路。该原型允许进行手持式固液萃取、无移液器的片上稀释,并将样品浓度调整到适当的分析动态工作范围内。一次性且可互连的均质器单元可在1分钟内从固体烘焙食品中萃取并基于三维筛网过滤过敏原蛋白。该均质器与一个3D打印的一体式芯片实验室(ULOC)微器件互连,该微器件用于将精确体积的样品提取物输送到试剂储存器中。试剂储存器用于在芯片上存储碳纳米颗粒标记的抗体和用于稀释的运行缓冲液。该手持式原型能够对固体样品进行完全均质化、固液蛋白质萃取、基于3D打印筛网的过滤、基于ULOC的稀释、混合、运输,并通过智能手机对固体烘焙食品中的榛子和花生过敏原进行检测,操作复杂度有限。多重侧向流动免疫分析(LFIA)能够在实际烘焙食品中检测低至0.1 ppm的过敏原,并且该系统已经可供消费者操作,证明了其在未来公民科学方法中的潜力。只要有LFIA可用,所设计的系统适用于食品安全之外的广泛分析应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/87fc/7567528/11eeec04442d/gr1_lrg.jpg

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