Introducing charge tag via click reaction in living cells for single cell mass spectrometry,Chemical Science

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Introducing charge tag via click reaction in living cells for single cell mass spectrometry
Chemical Science ( IF 7.6 ) Pub Date : 2020-06-23 , DOI: 10.1039/d0sc00259c
Meihui Zhuang ₁ , Zhuanghao Hou ₁ , Peiyao Chen ₁ , Gaolin Liang ₁ , Guangming Huang _{1,

2}

Affiliation

For single living cell mass spectrometry measurement, sensitivity is of great significance due to the extremely complicated chemical components of the cytoplasm. Higher sensitivity is always highly desired, especially for chemicals with low concentrations or poor mass spectrometry responses. Here, a quaternary ammonium salt group-based charge tag was designed to enhance the analytical performance for cysteine within single cells using induced nanoelectrospray mass spectrometry. While the charge tag was coupled to the analyte via biocompatible click reaction, viability of the living cells was maintained during in situ derivatization and following analysis. Enhanced sensitivity under physiological conditions for cysteine, at pH 7.4 and with highly concentrated salts, was achieved due to higher ionization efficiency of the charge tag. Therefore, the cysteine levels in single living HeLa cells and HepG2 cells were found to be in the range of 62.0 ± 3.4 μM and 49.6 ± 7.2 μM, respectively. Furthermore, the low cysteine levels in living single HeLa cells could be monitored, in the presence of cystine transporter inhibitor. Thus, this method provides a general strategy for in situ chemical derivatization for signal amplification in the field of single cell mass spectrometry.

中文翻译：

通过活细胞中的点击反应引入电荷标签用于单细胞质谱分析

对于单活细胞质谱测量来说，由于细胞质的化学成分极其复杂，灵敏度具有重要意义。始终高度期望更高的灵敏度，特别是对于浓度低或质谱响应差的化学品。在这里，设计了基于季铵盐基团的电荷标签，以提高使用诱导纳电喷雾质谱法对单细胞内半胱氨酸的分析性能。虽然电荷标签通过生物相容性点击反应与分析物偶联，但在原位衍生化和后续分析过程中活细胞的活力得以维持。由于电荷标签的电离效率较高，在生理条件下（pH 7.4 和高浓度盐），半胱氨酸的灵敏度得到了增强。因此，单个活 HeLa 细胞和 HepG2 细胞中的半胱氨酸水平分别在 62.0 ± 3.4 μM 和 49.6 ± 7.2 μM 范围内。此外，在存在胱氨酸转运蛋白抑制剂的情况下，可以监测活的单个 HeLa 细胞中的低半胱氨酸水平。因此，该方法为单细胞质谱领域中信号放大的原位化学衍生化提供了通用策略。

更新日期：2020-07-22

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