Development and Evaluation of a LC/UV/MS Peptide Mapping System

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Paul Rainville, Da Ren, Himanshu Gadgil, and Jeff Mazzeo[Waters]
Pittcon 2004
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Bradykinin, Human Serum Albumin, IgG1 tryptic digest
BioSuite PA-A C18 3µm Peptide Analysis Column 2.1 mm 150 mm
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Peptide mapping has been a conerstone assay in the characterization and quality control of biotherapeutics. Historically this method has been based on UV detection using TFA in the mobile phase. Recently, peptide mapping has become interfacted with mass spectrometry even in quality control. We developed a highly robust, reproducible new peptide mapping system that is designed both for UV and Mass Spectrometry detection. It couples a new chemistry using the mass spec freiendly mobile phase modifier, formic acid. Formic acid is requred for mass spectrometry compatiblility and eliminatse ion suppression and the subsequent loss of sensitivity by the mass spectrometer. Using a 3 hour antibody peptide map, we demonstrate the reproducibility and robustness in the identification and quantication of stability indicating modifications, oxidation, deamination, lysine varation etc

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