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Oral Sessions
Day 4, June 25（Wed.） 12:25-12:40
Room B (Maesato Center)
- 4B-O1-1225
Metal Ion-Enhanced ZIC-cHILIC StageTip for Simultaneous, Large-Scale Glycoproteomic and Phosphoproteomic Tissue Profiling in Breast Cancer
(1NTU/ Academia Sinica, 2Academia Sinica)
oHsiang-Chun Cheng1, Juanilita Waniwan2, Yu-Ju Chen2
Breast cancer, the most prevalent malignancy in women with > 2 million new cases annually, is a highly heterogeneous malignant disease with diverse clinical phenotypes. The membrane proteins modified by glycosylation and phosphorylation are critical for regulating cell-cell communication and signaling cascades. Despite large-scale tissue proteomics reveals patient heterogeneity, novel biomarker and therapeutic targets, simultaneous glycoproteomic and phosphoproteomic profiling in tissue is significantly under-explored. In this work, we utilized a metal ion-enhanced ZIC-cHILIC StageTip strategy (zwitterionic hydrophilic interaction chromatography with the exposed choline group) to simultaneously profile the glycoproteome and phosphoproteome in breast cancer tissue. To the best of our knowledge, this is the first Fe³⁺ in ZIC-cHILIC material which possess 79% specificity for phosphoproteomic analysis, with 3.5-fold coverage compared to Ga³⁺. Furthermore, we apply this strategy to paired of tumor and adjacent normal breast cancer tissue, enabling large-scale coverage of 17,270 GPs and 7,470 PPs from 200 μg tissue digest. Without protein immunoprecipitation, this strategy identified FDA-approved drug targets, such as EGFR, CD44, PD-L1, and integrin family members. This approach enables simultaneously profiling of both glycoproteome and phosphoproteome from a single sample. We aim to unravel the interplay between glycosylation and phosphorylation early recurrence luminal subtype.