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Poster Presentations
Day 4, June 25（Wed.）
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-AM-32（3C-O2-1440）
MiProChip: Microfluidic Device for Multiplexed Isotopic labeling-based Streamlined Single-cell Profiling
(1IoC, AS, 2NTU)
oHuan-Chi Chiu1,2, Tsai-Fang Chou1, Sofani Gebreyesus1, Guan-Fu Chen1, Hsiung-Lin Tu1, Yu-Ju Chen1,2
Microfluidic-based strategies provide flexibility and adaptability for bottom-up proteomics, making them ideal for single-cell analysis. In this study, we developed the Multiplexed Isotopic Labeling Integrated Proteomics Chip (MiProChip), which enhances multiplexing and sensitivity by integrating tandem mass tagging (TMT) and carrier-boosting strategies. MiProChip enables streamlined single-cell processing within a unified workflow, incorporating reaction chambers for single cells and 100-cell carriers. These chambers facilitate cell capture, lysis, protein reduction/alkylation, enzymatic digestion, TMT labeling, and desalting, preparing samples for mass spectrometry analysis. Using the TMT-10 plex model, MiProChip supports three modules, each with one carrier (100 cells) and nine single cells. When applied to lung cancer cells (PC9), it achieved 88–96% TMT labeling efficiency, identifying 3,322 peptides and 1,088 protein groups—4.7 times more than conventional methods. Functional module evaluation confirmed that the fully integrated setup yielded the highest protein identifications and Pearson correlation, demonstrating MiProChip's robustness. This is the first demonstration of TMT labeling within a microfluidic device for single-cell proteomics, underscoring its potential for large-scale clinical applications.