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首页> 外文期刊>Lab on a chip >Droplet CAR-Wash: continuous picoliter-scale immunocapture and washing
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Droplet CAR-Wash: continuous picoliter-scale immunocapture and washing

机译：液滴洗车：连续Picoliter级免疫敷贴和洗涤

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摘要

To address current limitations in adapting solid phase sample capture and washing techniques to continuously flowing droplet microfluidics, we have developed the "Coalesce-Attract-Resegment Wash" (CAR-Wash) approach. This module provides efficient, high-throughput magnetic washing by electrocoalescing magnetic bead-laden input droplets with a washing buffer flow and magnetophoretically transporting beads through the buffer into a secondary droplet formation streamline. In this work, we first characterized the technology in terms of throughput, sample retention, and flow-based exclusion of waste volume, demonstrating >500 Hz droplet processing with >98% bead retention and >100-fold dilution in final droplets. Next, we showed that the technique can be adapted to alternative commercially available magnetic beads with lower magnetite content per particle. Then, we demonstrated the CAR-Wash module's effectiveness in washing away a small molecule competitive inhibitor to restore the activity of magnetic bead-immobilized beta-galactosidase. Finally, we applied the system to immunomagnetically enrich a green fluorescent protein-histone H2B fusion protein from cell lysate while washing away mCherry and other lysate components. We believe this approach will bridge the gap between powerful biochemical and bioanalytical techniques and current droplet microfluidic capabilities, and we envision future application in droplet-based immunoassays, solid phase extraction, and other complex, multi-step operations.
机译：为了解决适应固体样品捕获和洗涤技术的当前限制，以持续流动的液滴微流体，我们开发了“Coalesce-Pressive-Recregment Wash”（洗车）方法。该模块通过电上磁珠 - 载带输入液滴提供有效，高通量的磁性洗涤，用洗涤缓冲液流流动并通过缓冲液将珠子通过缓冲液流入次级液滴形成流线线。在这项工作中，我们首先在废物的产量，样品保留和基于流量的排除方面表现了技术，展示了> 500Hz液滴加工> 98％珠子保留和最终液滴中的> 100倍稀释。接下来，我们表明该技术可以适用于可替代的市售磁珠，每个粒子具有下磁铁矿含量。然后，我们证明了洗车模块在洗涤小分子竞争性抑制剂时的有效性，以恢复磁珠固定的β-半乳糖苷酶的活性。最后，我们将该系统应用于免疫磁性，从细胞裂解物中富集绿色荧光蛋白组蛋白H2B融合蛋白，同时洗掉麦克里和其它裂解物组分。我们认为这种方法将弥合强大的生物化学和生物分析技术与电流液滴微流体能力之间的差距，并设想未来应用于基于液滴的免疫测定，固相提取和其他复杂的多步操作。

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