New Structure Mass Tag based on Zr-NMOF for Multiparameter and Sensitive Single-Cell Interrogating in Mass Cytometry
New Structure Mass Tag based on Zr-NMOF for Multiparameter and Sensitive Single-Cell Interrogating in Mass Cytometry
复制标题
基于 Ab 功能化 Zr-NMOF 的新型结构质量标签，用于质谱流式细胞术中的多参数和敏感单细胞生物标志物询问
DOI：
10.1002/adma.202008297
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发表时间：
2021-07-26
影响因子：
29.4
通讯作者：
Ding, Xianting
中科院分区：
文献类型：
--
作者：
Dang, Jingqi;Li, Hongxia;Ding, Xianting
质量细胞术，也称为飞行时间细胞术(CyTOF)，是一种新兴的强大的蛋白质组分析技术，它利用金属螯合聚合物(MCP)作为质量标签，同时在数百万个单独的细胞上询问高维生物标志物。然而，在典型的基于聚合物的质量标签系统下，其灵敏度和多重检测能力受到了很大的限制。本文报道了一种基于纳米金属有机骨架(NMOF)的新型结构质量标记物，用于细胞内多参数、灵敏的单细胞生物标记物的检测。采用调制剂/反应时间共调控法制备了尺寸均匀、携带10(5)个金属离子的单分散、胶体稳定、可在水中存放一年以上的纳米金属氧化物薄膜(ZRNMOF)(33 Nm)。在抗体的官能化过程中，锆质量标显示出特定的分子识别特性和与非靶细胞的最小交叉反应。此外，在小鼠脾细胞染色的多参数分析中，Zr质量标记与MCP质量标记兼容，该多参数分析利用了四个额外的通道，m/z=90、91、92、94，用于CyTOF中的单细胞免疫分析。与MCP质量标签相比，锆质标签提供了额外五倍的信号放大。这项工作为开发基于NMOF的质量标签用于CyTOF应用提供了基本的技术能力，为高维单细胞免疫图谱、低丰度抗原检测和开发新的条形码系统提供了可能性。
Mass cytometry, also called cytometry by time-of-flight (CyTOF), is an emerging powerful proteomic analysis technique that utilizes metal chelated polymer (MCP) as mass tags for interrogating high-dimensional biomarkers simultaneously on millions of individual cells. However, under the typical polymer-based mass tag system, the sensitivity and multiplexing detection ability has been highly restricted. Herein, a new structure mass tag based on a nanometal organic framework (NMOF) is reported for multiparameter and sensitive single-cell biomarker interrogating in CyTOF. A uniform-sized Zr-NMOF (33 nm) carrying 10(5) metal ions is synthesized under modulator/reaction time coregulation, which is monodispersed and colloidally stable in water for over one-year storage. On functionalization with an antibody, the Zr mass tag exhibits specific molecular recognition properties and minimal cross-reaction toward nontargeted cells. In addition, the Zr-mass tag is compatible with MCP mass tags in a multiparameter assay for mouse spleen cells staining, which exploits four additional channels, m/z = 90, 91, 92, 94, for single-cell immunoassays in CyTOF. Compared to the MCP mass tag, the Zr-mass tag provides an additional fivefold signal amplification. This work provides the fundamental technical capability for exploiting NMOF-based mass tags for CyTOF application, which opens up possibility of high-dimensional single-cell immune profiling, low abundant antigen detection, and development of new barcoding systems.