High-precision, automated peptide mapping of proteins

Instrument Type: UHPLC

The biopharmaceutical industry continues to develop protein-based biotherapeutics in increasing numbers. Due to their complexity and biotechnological production, there are many attributes that need to be analyzed to guarantee their safety and efficacy. Peptide mapping is used to measure several critical quality attributes (CQA) required for the characterization of any biotherapeutic protein. The analysis is used to confirm that the correct sequence has been expressed for the protein and to check for post-translational and chemical modifications.

SMART Digest compared to classic in-solution digestion of rituximab for in-depth peptide mapping characterization

Instrument Type: LCMS

This study compares the newly developed Thermo Scientific™ SMART Digest™ kit to classic in-solution protein digestion methods, focusing on protein sequence coverage and identified post-translational modifications (PTMs), including deamidation, oxidation, and glycosylation. A Thermo Scientific™ Acclaim™ VANQUISH™ C18 column with conventional water/acetonitrile-based gradients and the Thermo Scientific™ Vanquish™ Flex UHPLC system were used for separation in combination with the Thermo Scientific™ Q Exactive™ HF Hybrid Quadrupole-Orbitrap™ mass spectrometer.

Selecting buffers to remove uncertainty in tryptic digestion

Instrument Type: HPLC

The Thermo Scientific SMART Digest kit was used for the digestion of insulin. A variety of common buffer additives were compared to the SMART Digest buffer. Samples were analyzed on a Thermo Scientific Dionex UltiMate 3000 RSLC system via reversed-phase separation on a Thermo Scientific Accucore C18 column, with UV detection at 280 nm.