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Sample Preparation Method for Accurate Analysis of Nonenzymatic PTMs in Biotherapeutic Proteins with Peptide Mapping

Part # PS302

Abstract

Sergei Saveliev, Chris Hosfield, Mike Rosenblatt and Marjeta Urh
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI 53711

Non-enzymatic chemical modifications such as deamidation, disulfide bond scrambling and oxidation can affect the stability and efficacy of biotherapeutic proteins. Peptide mapping, the method of choice for monitoring these modifications, has a significant drawback which is that peptide mapping sample preparation induces the same modifications. 

We developed a sample preparation method which prevented the above side effects. In our method, all sample preparation steps including reduction, alkylation and digestion are performed at low pH (mildly acidic conditions).

Printed in USA.