A Method for Detecting Post Translational Modifiactions in Intact Proteins and Peptides

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Reb J. Russell II;Paul Rainville;Claude Mallet;Da Ren;Himanshu Gadgil [Waters]
HPLC 2003
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Atlantis dC18 3 µm Steel 4.6 mm 50 mm Atlantis dC18 3 µm Steel 2.1 mm 150 mm
Characterization of pharmaceutical proteins and peptides is an important analytical challenge that must be met in pharmaceutical and biotechnology companies today. Small chemical variations in proteins and peptides can be the difference between an active and an inactive therapeutic. In this study, we looked as some of the more common modifications; carbamylation, oxidation, formylation, deamidation, and acylation that are a byproduct of production, purification, and/or formulation. We have developed a comprehensive and rapid LC/MS methodology that can indicate the presence and degree of posttranslational modification using intact proteins or peptides. Our methodology is based on reversed-phase chromatography coupled with mass spectrometry since LC-MS is becoming a choice for the determination of post translation modifications.

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