Glycoproteome of Human lung cancer cell by LC-MSMS

We demonstrate the utility of lectin-magnetic nanoprobe coupled with Orbitrap HCD-CID-MS/MS for complementary glycotope-specific enrichment and site-specific glycosylation analysis of the glycoproteome. Using non-small cell lung cancer (NSCLC) cells as model, this approach allowed us to compare the glycosylation profiles of the drug resistant cell, PC9, and its resistant derivative, PC9-IR. The complementary enrichment allowed broader targeting of different glycoforms; using Byonic software, confident identification of a total of 2767 and 2290 non-redundant intact glycopeptides was achieved for PC9-IR and PC9, respectively. This also enabled glycan profiling of the onco-protein EGFR, without immunoprecipitation. Moreover, the glycan specificity afforded by the lectin@MNP allowed as to observe increased terminal and core fucosylation in EGFR from PC9-IR, which demonstrates the potential for protein glycosylation alterations in the resistant cell line. The multi-lectin nanoprobe-based affinity mass spectrometry presents a new method for large-scale glycoproteomic profiling of different sample types without prior fractionation or immunoprecipitation.