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A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells

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Abstract

To study the heterogeneity of target membrane proteins in single cells with cellular integrity, we proposed a simple and low-cost method to obtain the copy number of the membrane proteins. HeLa cells were labeled by FITC affinity bodies specifically targeting HER2 membrane proteins. The immunolabeled HeLa cells were quantified by a laboratory-built laser induced fluorescence detector. A series of fluorescent microspheres with known number of FITC molecules on the surface were used to establish the calibration curve, instead of the standard fluorescent solutions, because the morphology of the microspheres was similar to the cells, and the distribution of FITC on the spheres were similar to the distribution of HER2 on the HeLa. The fluorescence intensity of the cells was converted to the molecule number of HER2 by the calibration curve. A capillary electrophoresis system was used to drive the microspheres and cells through the detection window. The copy number of HER2 in HeLa cells ranged from 4,036 to 1,224,920 ± 100 (2.5–97.5%), and the median of copy numbers were 104,438 ± 100 per cell. This method for measuring low-abundance membrane proteins can be utilized for the initial exploration of proteomics in ordinary laboratories.

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Data Availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

Thanks for professor Liming Wang of Dalian Medical University for the supplement of cells. Thanks for associated professor Shenghong Li of Dalian Institute of Chemical Physics for her effective suggestions.

Funding

This research was funded by the Youth Innovation Promotion Association of the Chinese Academy of Sciences, grant number 2019186, and the Natural Science Foundation of China, grant number 22106014.

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Authors and Affiliations

  1. Department of Instrumentation & Analytical Chemistry, CAS Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, 116023, China

    Jiamin Li, Xuhui Geng & Yafeng Guan

  2. University of Chinese Academy of Sciences, Beijing, 100049, P.R. China

    Jiamin Li

  3. Key Laboratory of Deep-sea Composition Detection Technology of Liaoning Province, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, P. R. China

    Jiamin Li, Xuhui Geng & Yafeng Guan

  4. School of Chemistry and Chemical Engineering, Liaoning Normal University, Dalian, 116029, P.R. China

    Meng Shi

  5. Institute of Deep-Sea Science & Engineering, Chinese Academy of Sciences, 28 Luhuitou Road, Sanya, 572000, China

    Yafeng Guan

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  1. Jiamin Li
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Contributions

Jiamin Li and Meng Shi and established the analytical method on single cell analysis by CE-LIF system, and Jiamin Li wrote the main manuscript text. Xuhui Geng built the LIF. Yafeng Guan put forward the idea and guided the whole work.

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Correspondence to Yafeng Guan.

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Jiamin Li and Meng Shi are contributed equally.

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Li, J., Shi, M., Geng, X. et al. A Simple and Low-cost Preliminary Quantification of Target Membrane Protein in Single Cells. J Fluoresc (2023). https://doi.org/10.1007/s10895-023-03496-6

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