A simple,rapid,and practical method for single-cell proteomics based on mass-adaptive coating of synthetic peptides  

一种基于合成肽段涂层的单细胞蛋白质组学快速检测方法

作　　者：Siqi Li Kaichan Su Zhenkun Zhuang Qing Qin Lei Gao Yamei Deng Xuyang Liu Guixue Hou Longtao Wang Piliang Hao Huanming Yang Siqi Liu Hongming Zhu Yan Ren 李思奇;苏恺婵;庄镇堃;秦晴;高磊;邓亚美;刘旭阳;侯桂雪;王龙涛;郝丕良;杨焕明;刘斯奇;朱鸿明;任艳(BGI-Shenzhen,Shenzhen 528083,China;Department of Biology,University of Copenhagen,Copenhagen 2100,Denmark;BGI-Wuhan Clinical Laboratories,BGI-Shenzhen,Wuhan 430000,China;Department of Cardiology,Zhongshan Hospital,Fudan University,Shanghai Institute of Cardiovascular Disease,Shanghai 200032,China;Department of Cardiovascular Surgery,The First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China(USTC),Hefei 230001,China;School of Life Science and Technology,Shanghai Tech University,Shanghai 201210,China;Institute for Regenerative Medicine,Shanghai East Hospital,Tongji University School of Medicine,Shanghai 200120,China;Experiment Center for Science and Technology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]BGI-Shenzhen,Shenzhen 528083,China [2]Department of Biology,University of Copenhagen,Copenhagen 2100,Denmark [3]BGI-Wuhan Clinical Laboratories,BGI-Shenzhen,Wuhan 430000,China [4]Department of Cardiology,Zhongshan Hospital,Fudan University,Shanghai Institute of Cardiovascular Disease,Shanghai 200032,China [5]Department of Cardiovascular Surgery,The First Affiliated Hospital of USTC,Division of Life Sciences and Medicine,University of Science and Technology of China(USTC),Hefei 230001,China [6]School of Life Science and Technology,Shanghai Tech University,Shanghai 201210,China [7]Institute for Regenerative Medicine,Shanghai East Hospital,Tongji University School of Medicine,Shanghai 200120,China [8]Experiment Center for Science and Technology,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China

出　　处：《Science Bulletin》2022年第6期581-584,M0003,共5页科学通报（英文版）

基　　金：supported by the National Key R&D Program of China(2020YFE0202200);the National Natural Science Foundation of China(82070365);the Basic Research General Program of Shenzhen(JCYJ20190807103407873)。

摘　　要：Since a typical mammalian cell contains only 200–300 picograms of proteins and the lack of a proteome amplification strategy[1],a practical and high-throughput approach for single-cell proteomics has long been awaited.A series of methods have been explored to make single-cell proteomics available.The design of nanoPOTS and OAD chip-based processes minimizes sample preparation steps and volumes to reduce sample loss[2,3].单细胞层面的蛋白质组学研究极具挑战.本文报道了一种简单、快速的单细胞蛋白质组学方法(Mad-CASP),创新地设计了一套质谱兼容的合成肽段:(1)样品制备过程中,合成肽段的包被可以有效地减少样品损失;(2)质谱数据采集时,合成肽段的信号被摒除,对样品信号无影响;(3)全流程无需特殊设备,可在3~4小时内完成96孔样品的高通量制备.结合图谱库和新的质谱数据采集模式,Mad-CASP技术可从单个HeLa细胞中鉴定出124093个蛋白质.随后利用Mad-CASP技术分析了来自健康人和冠状动脉慢性完全闭塞(CTO)患者的240个外周血CD34^(+)单细胞.基于单细胞蛋白图谱,本研究首次揭示外周血CD34^(+)细胞的功能亚型,证实其在CTO进展中的“双刃剑”作用,提示靶向乙醛脱氢酶2在CTO治疗中的潜在作用.综上,本研究利用Mad-CASP技术首次解析了循环CD34^(+)细胞的单细胞蛋白质图谱,提示Mad-CASP技术在稀缺临床样本的单细胞蛋白质组学检测方面极具潜力.

关 键 词：乙醛脱氢酶2 单细胞蛋白质 蛋白质组学 潜在作用 质谱数据 数据采集模式 HELA细胞 快速检测方法 

分 类 号：Q26[生物学—细胞生物学]