Introducing charge tag via click reaction in living cells for single cell mass spectrometry
Chemical Science 11(28):7308-7312 Jul 2020 

Zhuang, Meihui; Hou, Zhuanghao; Chen, Peiyao; Liang, Gaolin; Huang, Guangming


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. 

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