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.

To demonstrate the use of a multi-pump UHPLC system and enable tandem analysis with two columns in parallel, addressing productivity and throughput improvement of existing LC-MS methods.

The Thermo Scientific SMART Digest Kit is applied for generation of peptides from therapeutic monoclonal antibodies in 60 minutes. Peptide mapping samples were analysed on a Thermo Scientific UltiMate U3000 RSLC system coupled to a Thermo Scientific Q Exactive HF with Biopharma Option. Separation of sample peptides was performed on a Thermo Scientific Acclaim RSLC 120, C18, 2.2 µm analytical column using a 30 minute separation gradient, yielding excellent chromatographic separation and high sequence coverage.

We present a summary of a sensitive and robust HRAM LC-MS method using a Thermo Scientific Q Exactive mass spectrometer for the identification and detection of marker proteins in raw meat samples. Four proteotypic peptides were used to perform accurate meat speciation and authenticity. Myoglobin tryptic peptides from each species were detected with an observed m/z below 1.3 ppm. This targeted method allowed for the detection of undesired meat species down to 1% (w/w) of the entire sample.

Peptide mapping is one of the major analyses in biotherapeutics characterization. Here we show peptide mapping with the Thermo Scientific Vanquish UHPLC system. An efficient and reliable separation of peptides obtained from a bovine serum albumin (BSA) digest is done on a Thermo Scientific Acclaim RSLC 120, C18, 2.2 µm Analytical column. Excellent retention time and area precisions as well as peak capacities were achieved over a time period of more than 80 injections.