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Rapid microfluidic platform for screening and enrichment of cells secreting virus neutralizing antibodies
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-06-13 , DOI: 10.1039/d2lc00018k Weikang Nicholas Lin 1 , Matthew Zirui Tay 2 , Joel Xu En Wong 2 , Chia Yin Lee 3 , Siew-Wai Fong 2 , Cheng-I Wang 3 , Lisa Fong Poh Ng 2, 4, 5, 6 , Laurent Renia 2, 7, 8 , Chia-Hung Chen 8 , Lih Feng Cheow 1, 9, 10
Lab on a Chip ( IF 6.1 ) Pub Date : 2022-06-13 , DOI: 10.1039/d2lc00018k Weikang Nicholas Lin 1 , Matthew Zirui Tay 2 , Joel Xu En Wong 2 , Chia Yin Lee 3 , Siew-Wai Fong 2 , Cheng-I Wang 3 , Lisa Fong Poh Ng 2, 4, 5, 6 , Laurent Renia 2, 7, 8 , Chia-Hung Chen 8 , Lih Feng Cheow 1, 9, 10
Affiliation
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As part of the body's immune response, antibodies (Abs) have the ability to neutralize pathogenic viruses to prevent infection. To screen for neutralizing Abs (nAbs) from the immune repertoire, multiple screening techniques have been developed. However, conventional methods have a trade-off between screening throughput and the ability to screen for nAbs via their functional efficacy. Although droplet microfluidic platforms have the ability to bridge this disparity, the majority of such reported platforms still rely on Ab-binding assays as a proxy for function, which results in irrelevant hits. Herein, we report the multi-module Droplet-based Platform for Effective Antibody RetrievaL (DROP-PEARL) platform, which can achieve high-throughput enrichment of Ab-secreting cells (ASCs) based on the neutralizing activity of secreted nAbs against the a target virus. In this study, in-droplet Chikungunya virus (CHIKV) infection of host cells and neutralization was demonstrated via sequential delivery of viruses and host cells via picoinjection. In addition, we demonstrate the ability of the sorting system to accurately discriminate and isolate uninfected droplets from a mixed population of droplets at a rate of 150 000 cells per hour. As a proof of concept, a single-cell neutralization assay was performed on two populations of cells (nAb-producing and non-Ab producing cells), and up to 2.75-fold enrichment of ASCs was demonstrated. Finally, we demonstrated that DROP-PEARL is able to achieve similar enrichment for low frequency (∼2%) functional nAb-producing cells in a background of excess cells secreting irrelevant antibodies, highlighting its potential prospect as a first round enrichment platform for functional ASCs. We envision that the DROP-PEARL platform could potentially be used to accelerate the discovery of nAbs against other pathogenic viral targets, and we believe it will be a useful in the ongoing fight against biological threats.
中文翻译：
用于筛选和富集分泌病毒中和抗体的细胞的快速微流控平台
作为身体免疫反应的一部分，抗体 (Abs) 具有中和致病病毒以预防感染的能力。为了从免疫组库中筛选中和抗体 (nAb)，已经开发了多种筛选技术。然而，传统方法在筛选通量和筛选 nAb 的能力之间需要权衡取舍。它们的功能功效。尽管液滴微流体平台有能力弥合这种差异，但大多数此类报告的平台仍然依赖于抗体结合测定作为功能的代理，这会导致不相关的命中。在此，我们报告了基于液滴的多模块有效抗体检索平台 (DROP-PEARL) 平台，该平台可以基于分泌的 nAb 对靶标的中和活性实现 Ab 分泌细胞 (ASCs) 的高通量富集病毒。在这项研究中，通过连续递送病毒和宿主细胞，证明了宿主细胞的液滴内基孔肯雅病毒 (CHIKV) 感染和中和作用皮内注射。此外，我们展示了分拣系统以每小时 150 000 个细胞的速度从混合液滴群体中准确区分和分离未感染液滴的能力。作为概念验证，对两个细胞群（产生 nAb 的细胞和不产生 Ab 的细胞）进行了单细胞中和测定，并证明了 ASC 的富集度高达 2.75 倍。最后，我们证明了在过量细胞分泌无关抗体的背景下，DROP-PEARL 能够对低频（~2%）功能性 nAb 产生细胞实现类似的富集，突出了其作为功能性 ASC 第一轮富集平台的潜在前景. 我们设想 DROP-PEARL 平台可能用于加速发现针对其他致病病毒靶点的 nAb，
更新日期：2022-06-13
中文翻译：
用于筛选和富集分泌病毒中和抗体的细胞的快速微流控平台
作为身体免疫反应的一部分，抗体 (Abs) 具有中和致病病毒以预防感染的能力。为了从免疫组库中筛选中和抗体 (nAb)，已经开发了多种筛选技术。然而，传统方法在筛选通量和筛选 nAb 的能力之间需要权衡取舍。它们的功能功效。尽管液滴微流体平台有能力弥合这种差异，但大多数此类报告的平台仍然依赖于抗体结合测定作为功能的代理，这会导致不相关的命中。在此，我们报告了基于液滴的多模块有效抗体检索平台 (DROP-PEARL) 平台，该平台可以基于分泌的 nAb 对靶标的中和活性实现 Ab 分泌细胞 (ASCs) 的高通量富集病毒。在这项研究中，通过连续递送病毒和宿主细胞，证明了宿主细胞的液滴内基孔肯雅病毒 (CHIKV) 感染和中和作用皮内注射。此外，我们展示了分拣系统以每小时 150 000 个细胞的速度从混合液滴群体中准确区分和分离未感染液滴的能力。作为概念验证，对两个细胞群（产生 nAb 的细胞和不产生 Ab 的细胞）进行了单细胞中和测定，并证明了 ASC 的富集度高达 2.75 倍。最后，我们证明了在过量细胞分泌无关抗体的背景下，DROP-PEARL 能够对低频（~2%）功能性 nAb 产生细胞实现类似的富集，突出了其作为功能性 ASC 第一轮富集平台的潜在前景. 我们设想 DROP-PEARL 平台可能用于加速发现针对其他致病病毒靶点的 nAb，
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