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单域抗体功能化流动池上细胞的快速捕获与标记

Rapid capture and labeling of cells on single domain antibodies-functionalized flow cell.

作者信息

Chen Guan-Yu, Li Zeyang, Duarte Joao N, Esteban Alexandre, Cheloha Ross W, Theile Christopher S, Fink Gerald R, Ploegh Hidde L

机构信息

Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Institute of Biomedical Engineering, National Chiao Tung University, Hsinchu 30010, Taiwan; Department of Biological Science and Technology, National Chiao Tung University, Hsinchu 30010, Taiwan.

Whitehead Institute for Biomedical Research, Cambridge, MA, USA.

出版信息

Biosens Bioelectron. 2017 Mar 15;89(Pt 2):789-794. doi: 10.1016/j.bios.2016.10.015. Epub 2016 Oct 5.

Abstract

Current techniques to characterize leukocyte subgroups in blood require long sample preparation times and sizable sample volumes. A simplified method for leukocyte characterization using smaller blood volumes would thus be useful in diagnostic settings. Here we describe a flow system comprised of two functionalized graphene oxide (GO) surfaces that allow the capture of distinct leukocyte populations from small volumes blood using camelid single-domain antibodyfragments (VHHs) as capture agents. We used site-specifically labeled leukocytes to detect and identify cells exposed to fungal challenge. Combining the chemical and optical properties of GO with the versatility of the VHH scaffold in the context of a flow system provides a quick and efficient method for the capture and characterization of functional leukocytes.

摘要

目前用于血液中白细胞亚群特征分析的技术需要较长的样本制备时间和大量的样本体积。因此,一种使用较小血量进行白细胞特征分析的简化方法在诊断环境中会很有用。在这里,我们描述了一种流动系统,该系统由两个功能化的氧化石墨烯(GO)表面组成,允许使用骆驼科单域抗体片段(VHHs)作为捕获剂从小体积血液中捕获不同的白细胞群体。我们使用位点特异性标记的白细胞来检测和识别受到真菌攻击的细胞。在流动系统的背景下,将GO的化学和光学特性与VHH支架的多功能性相结合,为功能性白细胞的捕获和特征分析提供了一种快速有效的方法。

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