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通过低信号损失的单细胞西部斑块与光定的阿普塔米尔进行多重蛋白质分析

Li Xu^1,2, Haiyang Xie¹, Boqian Wang¹

¹Department of Anesthesiology and Surgical Intensive Care Unit, Xinhua Hospital, School of Medicine and School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200230, China.

Analytical chemistry

|July 17, 2023

概括

我们开发了光灭的基于Aptamer的单细胞西部斑点 (FAS-WB),以克服多重蛋白检测中的信号损失. 这种新方法使单细胞中高效,成本效益和高分辨率的蛋白质分析成为可能.

科学领域:

生物化学生物化学
分子生物学分子生物学
蛋白质组学是指蛋白质组学.

背景情况:

单细胞西部涂抹 (scWB) 提供高分辨率蛋白质分析,但在抗体剥离过程中遭受信号损失,限制多重复合.
在单细胞中检测多重蛋白质对于理解细胞异质性和功能至关重要.

研究的目的:

引入基于Aptamer的单细胞西部斑点 (FAS-WB) 以光灭,以在单细胞分辨率下检测多重蛋白质.
为了解决与传统基于抗体的scWB相关的信号损失和成本的局限性.

主要方法:

开发了FAS-WB,使用最小尺寸的aptamer探头来实现快速的内凝动态.
采用光标记的阿普坦,与补充火线协调,以防止在化过程中信号损失.
使用FAS-WB.证明了四种生物标记蛋白 (EpCAM,PTK7,HER2,CA125) 的联合成像.

主要成果:

与传统scWB相比,FAS-WB在单细胞分辨率下实现了多重蛋白质检测,信号损失明显较低.
该方法证明了信号与噪声比的增强.
FAS-WB节省了时间 (每周期不到25分钟),并且具有成本效益 (抗体探针成本的1000分之一).

结论:

FAS-WB提供了一个高效,经济和节省时间的平台,以单细胞分辨率进行多重蛋白质分析.
基于aptamer的方法克服了传统scWB的关键局限性,使其在细胞生物学研究中具有更广泛的应用.

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