On-Chip Analysis of Protein Secretion from Single Cells Using Microbead Biosensors,ACS Sensors

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On-Chip Analysis of Protein Secretion from Single Cells Using Microbead Biosensors
ACS Sensors ( IF 8.2 ) Pub Date : 2023-01-29 , DOI: 10.1021/acssensors.2c02148
Diana F Cedillo-Alcantar ₁ , Roberto Rodriguez-Moncayo ₁ , Jose L Maravillas-Montero ₂ , Jose L Garcia-Cordero _{1,

3}

Affiliation

The profiling of the effector functions of single immune cells─including cytokine secretion─can lead to a deeper understanding of how the immune system operates and to potential diagnostics and therapeutical applications. Here, we report a microfluidic device that pairs single cells and antibody-functionalized microbeads in hydrodynamic traps to quantitate cytokine secretion. The device contains 1008 microchambers, each with a volume of ∼500 pL, divided into six different sections individually addressed to deliver an equal number of chemical stimuli. Integrating microvalves allowed us to isolate cell/bead pairs, preventing cross-contamination with factors secreted by adjacent cells. We implemented a fluorescence sandwich immunoassay on the biosensing microbeads with a limit of detection of 9 pg/mL and were able to detect interleukin-8 (IL-8) secreted by single blood-derived human monocytes in response to different concentrations of LPS. Finally, our platform allowed us to observe a significant decrease in the number of IL-8-secreting monocytes when paracrine signaling becomes disrupted. Overall, our platform could have a variety of applications for which the analysis of cellular function heterogeneity is necessary, such as cancer research, antibody discovery, or rare cell screening.

中文翻译：

使用微珠生物传感器对单细胞蛋白质分泌进行片上分析

分析单个免疫细胞的效应子功能（包括细胞因子分泌）可以加深对免疫系统运作方式的了解，以及潜在的诊断和治疗应用。在这里，我们报告了一种微流体装置，该装置将单细胞和抗体功能化微珠配对在流体动力学陷阱中以定量细胞因子分泌。该装置包含 1008 个微室，每个微室的体积约为 500 pL，分为六个不同的部分，分别处理以提供相同数量的化学刺激。集成微型阀使我们能够分离细胞/珠子对，防止与相邻细胞分泌的因子交叉污染。我们对生物传感微珠实施了荧光夹心免疫测定，检测限为 9 pg/mL，并且能够检测单个血源性人单核细胞分泌的白细胞介素 8 (IL-8) 以响应不同浓度的 LPS。最后，我们的平台使我们能够观察到当旁分泌信号中断时分泌 IL-8 的单核细胞数量显着减少。总的来说，我们的平台可以有多种需要分析细胞功能异质性的应用，例如癌症研究、抗体发现或稀有细胞筛选。

更新日期：2023-01-29

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