Pressurized Online Pepsin Digestion of mAb IgG2 for Hydrogen Deuterium Exchange Mass Spectrometry

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Jing Fang, Ying Qing Yu, and Weibin Chen
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Application Notes
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Application Notes
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SYNAPT G2-Si High Definition Mass Spectrometry

Here we report an improvement in protein digestion efficiency using a high-pressure, sustainable pepsin column – the Enzymate™ Digestion Column.

An enhanced-pressure (up to ~ 15,000 psi), sustainable BEH column with immobilized pepsin has been successfully applied to the Waters® ACQUITY UPLC® M-Class System with HDX Technology. Online peptic digestion of the monoclonal antibody (mAb) IgG2 led to increased digestion efficiency using high pressure (HP) when compared to ambient or normal pressure (NP).

High-pressure digestion yielded an increased number of overlapping peptides with shorter lengths, which resulted in greater protein sequence coverage and higher spatial resolution compared to normal-pressure digestion.

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