Improved Deglycosylation of N-linked Glycoproteins and Sample Preparations for MALDI-TOF MS Analysis

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Ying Qing Yu, Weibin Chen, Martin Gilar and John C. Gebler [Waters]
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N-Linked Glycoproteins
An acid labile surfactant (ALS) was used to replace SDS for denaturing N-linked glycoprotein prior to enzymatic deglycosylation reactions. This anionic surfactant was shown previously to denature proteins without inhibiting the activity of enzymes (Anal. Chem. 75, 6023-28, 2003). ALS was easily decomposed via direct acidification on a MALDI target; therefore minimizing the subsequent sample preparation for MALDITOF MS analysis of oligosaccharides released by PNGase F. The MALDI target used has a chemically modified hydrophobic surface which allows the extraction of oligosaccharides with pure water from the deglycosylated proteins. Our method requires less time for deglycosylation reaction and minimizes sample clean up procedures. The hydrophobic MALDI target enables the separation of oligosaccharides from proteins, therefore significantly improveing their ion signals.

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