Abstract
Low-input proteomics, which treats tens to hundreds of mammalian cells, is the gap between standard proteomics and single-cell proteomics. Low-input proteomics is widely applicable and needs special sample preparation methods to achieve deep proteome profiling. This chapter describes protocols for the preparation and application of an easy-to-use and scalable device for processing low-input samples. Protein preconcentration, impurity removal, reduction, alkylation, digestion, and desalting are fully integrated into this workflow, and the device can be directly connected to online nanoLC-MS to avoid sample transfer.
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Acknowledgments
This work is supported by grants from the China State Key Basic Research Program Grants (2021YFA1301601, 2021YFA1301602, 2021YFA1302603, 2020YFE0202200, and 2022YFC3401104), the National Natural Science Foundation of China (22125403, 22074060, 22150610470, and 92253304), the Shenzhen Innovation of Science and Technology Commission (JCYJ20200109140814408, JCYJ20210324120210029, and JCYJ20200109141212325), and Guangdong province (2019B151502050).
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Yang, Y., Tian, R. (2024). Fully Integrated Online Strategy for Highly Sensitive Proteome Profiling. In: Vegvari, A., Teppo, J., Zubarev, R.A. (eds) Mass Spectrometry Based Single Cell Proteomics. Methods in Molecular Biology, vol 2817. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3934-4_6
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DOI: https://doi.org/10.1007/978-1-0716-3934-4_6
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3933-7
Online ISBN: 978-1-0716-3934-4
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