Solid Phase Extraction of N‑Linked Glycopeptides
Using Hydrazide Tip
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Abstract
Glycoproteome contains
valuable information where biomarkers may
be discovered for disease diagnosis and monitoring. Nowadays, with
the ever-increasing performances of mass spectrometers, the emphasis
is shifting to the sample preparation for better throughput and reproducibility.
Therefore, to facilitate high throughput N-linked glycopeptide isolation,
in this study, a novel hydrazide tip was devised and an integrated
workflow of N-linked glycopeptide isolation using hydrazide tips was
presented. With the use of bovine fetuin as a standard glycoprotein,
the incubation time was determined for each major step of glycopeptide
isolation. With the use of commercially available human serum, multiple
parallel isolations of glycopeptides were performed using hydrazide
tips with a liquid handling robotic system. We demonstrated that,
with the hydrazide tips, the processing time was significantly decreased
from 3 to 4 days to less than 8 h with excellent reproducibility.
The hydrazide pipet tips have great potential in achieving automation
of N-linked glycopeptide isolation for high-throughput sample preparation
when used in combination with liquid handling robotic systems