Localized cell-surface sampling of a secreted factor using cell-targeting beads.,Analytical Chemistry

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Localized cell-surface sampling of a secreted factor using cell-targeting beads.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-09-17 , DOI: 10.1021/acs.analchem.0c02578
Tammi L van Neel ₁ , Samuel B Berry ₁ , Erwin Berthier ₁ , Ashleigh B Theberge _{1,

2}

Affiliation

Intercellular communication through the secretion of soluble factors plays a vital role in a wide range of biological processes (e.g., homeostasis, immune response), yet identification and quantification of many of these factors can be challenging due to their degradation or sequestration in cell culture media prior to analysis. Here, we present a customizable bead-based system capable of simultaneously binding to live cells (through antibody-mediated cell tethering) and capturing cell-secreted molecules. Our functionalized beads capture secreted molecules (e.g., hepatocyte growth factor secreted by fibroblasts) that are diminished when sampled via traditional supernatant analysis techniques (p < 0.05), effectively rescuing a reduced signal in the presence of neutralizing components in the cell culture media. Our system enables capture and analysis of molecules integral to chemical communication that would otherwise be markedly decreased prior to analysis.

中文翻译：

使用细胞靶向珠对分泌因子进行局部细胞表面采样。

通过可溶性因子的分泌进行的细胞间通讯在广泛的生物过程（例如稳态、免疫反应）中起着至关重要的作用，但由于它们在细胞培养基中的降解或隔离，许多这些因子的鉴定和量化可能具有挑战性在分析之前。在这里，我们提出了一种可定制的基于珠子的系统，能够同时与活细胞结合（通过抗体介导的细胞系留）并捕获细胞分泌的分子。我们的功能化微珠捕获分泌分子（例如，成纤维细胞分泌的肝细胞生长因子），当通过传统上清液分析技术采样时，这些分子会减少（p< 0.05)，在细胞培养基中存在中和成分的情况下有效地挽救降低的信号。我们的系统能够捕获和分析化学通讯不可或缺的分子，否则这些分子在分析之前会显着减少。

更新日期：2020-10-21

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