High-throughput secretomic analysis of single cells to assess functional cellular heterogeneity.
High-throughput secretomic analysis of single cells to assess functional cellular heterogeneity.
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DOI：
10.1021/ac400082e
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发表时间：
2013-02-19
影响因子：
7.4
通讯作者：
Fan, Rong
中科院分区：
文献类型：
--
作者：
Lu, Yao;Chen, Jonathan J.;Mu, Luye;Xue, Qiong;Wu, Yu;Wu, Pei-Hsun;Li, Jie;Vortmeyer, Alexander O.;Miller-Jensen, Kathryn;Wirtz, Denis;Fan, Rong
分泌的蛋白质决定了人类健康和疾病中的一系列细胞功能。由于高度的细胞异质性，更重要的是，单个细胞的多功能性，同时测量单个细胞中的一系列蛋白质并快速并行地快速分析大量单细胞（超过 1000 个）的需求尚未得到满足。我们描述了一个简单的生物分析测定平台，该平台由大量亚纳升微室组成，与高密度抗体条形码微阵列集成，用于并行检测一千多个单细胞的高度多重蛋白质。该平台已经针对细胞系和复杂的生物样品（例如来自患者的原代细胞）进行了测试。我们观察到单细胞分泌特征之间存在明显的异质性，这些异质性首次可以与细胞的物理行为（例如迁移）直接相关。与最先进的蛋白质分泌测定（例如 ELISpot 和新兴的微技术测定）相比，我们的方法提供高通量和高多样性。它还具有许多临床医生友好的功能，例如易于操作、低样本消耗和标准化数据分析，代表了一种潜在的变革性工具，可以对患者的细胞功能和免疫力进行信息监测。
Secreted proteins dictate a range of cellular functions in human health and disease. Due to the high degree of cellular heterogeneity and, more importantly, polyfunctionality of individual cells, there is an unmet need to simultaneously measure an array of proteins from single cells and to rapidly assay a large number of single cells (more than 1000) in parallel. We describe a simple bioanalytical assay platform consisting of a large array of sub-nanoliter microchambers integrated with high-density antibody barcode microarrays for highly multiplexed protein detection from over a thousand single cells in parallel. This platform has been tested for both cell lines and complex biological samples such as primary cells from patients. We observed distinct heterogeneity among the single cell secretomic signatures that, for the first time, can be directly correlated to the cells’ physical behavior such as migration. Compared to the state-of-the-art protein secretion assay such as ELISpot and emerging microtechnology-enabled assays, our approach offers both high throughput and high multiplicity. It also has a number of clinician-friendly features such as ease of operation, low sample consumption and standardized data analysis, representing a potentially transformative tool for informative monitoring of cellular function and immunity in patients.
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影响因子：
48
作者：
Chen, Songming;LaRoche, Tom;Haab, Brian B.
通讯作者：
Haab, Brian B.
影响因子：
46.9
作者：
Love, J. Christopher;Ronan, Jehnna L.;Ploegh, Hidde L.
通讯作者：
Ploegh, Hidde L.
DOI：
10.1126/science.1198704
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bendall SC;Simonds EF;Qiu P;Amir el-AD;Krutzik PO;Finck R;Bruggner RV;Melamed R;Trejo A;Ornatsky OI;Balderas RS;Plevritis SK;Sachs K;Pe'er D;Tanner SD;Nolan GP
通讯作者：
Nolan GP
影响因子：
50.3
作者：
Kotecha N;Flores NJ;Irish JM;Simonds EF;Sakai DS;Archambeault S;Diaz-Flores E;Coram M;Shannon KM;Nolan GP;Loh ML
通讯作者：
Loh ML
DOI：
10.1158/1940-6207.capr-10-0234
发表时间：
2010-11
期刊：
Cancer prevention research (Philadelphia, Pa.)
影响因子：
--
作者：
Michor F;Polyak K
通讯作者：
Polyak K