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AutoGP 2: an improved fully automated high-throughput system for simultaneously enriching and separating intact glycopeptides

ABSTRACT: An improved fully automated high-throughput system, named as AutoGP 2, for simultaneously enriching and separating intact glycopeptides has been developed in this study by introducing a second 10-port valve (valve 2) to the previously developed AutoGP system with only a 10-port valve (valve 1). Specifically, the addition of the second 10-port valve in the new AutoGP 2 system allows simultaneous enrichment and separation of intact glycopeptides, which significantly improve the sample analysis throughput as compared to previously developed AutoGP system. The optimal operating parameters of the AutoGP 2 were systematically evaluated, and the intra-day and inter-day reproducibility of this system were assessed through 24-h and 15-day long stress tests. The experimental evaluation has shown outstanding sensitivity and reproducibility of AutoGP 2. Over 560 site-specific glycans were identified from 1 μg peptide mixture with median CV of 10.8% in triplicate sample analyses. By coupling the AutoGP 2 with a high resolution Orbitrap mass spectrometer, the duty cycle of the instrument for a single sample analysis was increased from 55% to 80%. Specifically, 166 minutes were saved for every two-sample analyses in the AutoGP2 as compared with the previous AutoGP system, which increased the sample analysis throughput by 1.5 times. All the experimental results have consistently shown that the AutoGP 2 is well-suited for large cohort clinical glycoproteomic sample analysis and very effective in the sample amount limited glycoproteomic analysis.

ORGANISM(S): Mus Musculus

SUBMITTER: Keqi Tang

PROVIDER: PXD069435 | iProX | Tue Oct 14 00:00:00 BST 2025

REPOSITORIES: iProX

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