Details

Autor(en) / Beteiligte

• Woo, Jongmin
• Williams, Sarah M.
• Markillie, Lye Meng
• Feng, Song
• Tsai, Chia-Feng
• Aguilera-Vazquez, Victor
• Sontag, Ryan L.
• Moore, Ronald J.
• Hu, Dehong
• Mehta, Hardeep S.
• Cantlon-Bruce, Joshua
• Liu, Tao
• Adkins, Joshua N.
• Smith, Richard D.
• Clair, Geremy C.
• Pasa-Tolic, Ljiljana
• Zhu, Ying

Titel

High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip

Ist Teil von

• Nature communications, 2021-10, Vol.12 (1), p.1-11, Article 6246

Ort / Verlag

London: Nature Publishing Group

Erscheinungsjahr

2021

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Abstract Global quantification of protein abundances in single cells could provide direct information on cellular phenotypes and complement transcriptomics measurements. However, single-cell proteomics is still immature and confronts many technical challenges. Herein we describe a nested nanoPOTS (N2) chip to improve protein recovery, operation robustness, and processing throughput for isobaric-labeling-based scProteomics workflow. The N2 chip reduces reaction volume to <30 nL and increases capacity to >240 single cells on a single microchip. The tandem mass tag (TMT) pooling step is simplified by adding a microliter droplet on the nested nanowells to combine labeled single-cell samples. In the analysis of ~100 individual cells from three different cell lines, we demonstrate that the N2 chip-based scProteomics platform can robustly quantify ~1500 proteins and reveal membrane protein markers. Our analyses also reveal low protein abundance variations, suggesting the single-cell proteome profiles are highly stable for the cells cultured under identical conditions.