Quantification of Intracellular Proteins in Single Cells Based on Engineered Picoliter Droplets,Langmuir

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Quantification of Intracellular Proteins in Single Cells Based on Engineered Picoliter Droplets
Langmuir ( IF 3.7 ) Pub Date : 2022-06-24 , DOI: 10.1021/acs.langmuir.2c00341
Weizhi Liu ₁ , Ruihua Zhang ₁ , Shanqing Huang ₁ , Xingrui Li ₁ , Wanling Liu ₁ , Jianhui Zhou ₁ , Lin Zhu ₁ , Yanling Song _{1,

2} , Chaoyong Yang _{1,

2}

Affiliation

Unlike conventional bulk measurements, single-cell protein analysis permits quantification of protein expression in individual cells. This has shed light on the cell-to-cell variation in heterogeneous biological systems, such as solid tumors, brain tissues, and developing embryos. Herein, a microfluidic method is developed to profile protein expression in individual cells by performing single-cell intracellular protein immunoassay in picoliter paired droplets. The high sensitivity of single-cell protein analysis on a chip is achieved by the confined reaction volume of picoliter droplets, efficient kinetic characteristics of the immunoassay through active mixing, and minimum single-cell protein loss by integrated operations. The abundance of an intracellular prostate specific antigen at the single-cell level is measured, and then the platform is applied to identify cell types and investigate heterogeneity within cell populations. Overall, a paired chip for single-cell immunoassay establishes a foundation for parallel, sensitive, and integrated protein quantification at the single-cell level and will find wide applications in the field of single-cell proteomics.

中文翻译：

基于工程皮升液滴的单细胞胞内蛋白定量

与传统的批量测量不同，单细胞蛋白质分析允许对单个细胞中的蛋白质表达进行量化。这揭示了异质生物系统中的细胞间变异，例如实体瘤、脑组织和发育中的胚胎。在此，开发了一种微流体方法，通过在皮升配对液滴中进行单细胞细胞内蛋白质免疫测定来分析单个细胞中的蛋白质表达。芯片上单细胞蛋白分析的高灵敏度是通过皮升液滴的有限反应体积、通过主动混合实现免疫测定的高效动力学特性以及通过集成操作将单细胞蛋白损失最小化来实现的。测量单细胞水平的细胞内前列腺特异性抗原的丰度，然后将该平台用于识别细胞类型并研究细胞群内的异质性。总体而言，用于单细胞免疫测定的配对芯片为单细胞水平的并行、灵敏和集成蛋白质定量奠定了基础，并将在单细胞蛋白质组学领域得到广泛应用。

更新日期：2022-06-24

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