This application shows the quantification of multiple impurities with a single calibrant by taking advantage of the uniform response of the charged aerosol detector (CAD). The inverse gradient compensation wizard is also highlighted.
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.
Peptide mapping is one of the major analyses in biotherapeutics characterization. Here we show peptide mapping with the Thermo Scientific Vanquish Flex UHPLC system. An efficient and reliable separation of peptides obtained from a bovine serum albumin (BSA) digest is done on an Thermos Scientific Acclaim RSLC 120, C18, 2.2 µm Analytical column. Excellent retention time and area precisions allow reliable qualitative and quantitative data interpretation.
The Thermo Scientific Vanquish UHPLC system was used in combination with the Thermo Scientific Acclaim RSLC 120 C18, 2.2 µm, 2.1x250 mm column to provide high resolution peptide separation of a monoclonal antibody digest. The column was thermostatted at 40, 60, and 80 °C. Both still-air and forced-air mode were used. The highest peak capacity was obtained at 80 °C, and with still-air thermostatting mode. At 40 °C, the peak capacity obtained with still-air and forced-air was comparable.
A sample of a monoclonal antibody, was separated by a Thermo Scientific UltiMate 3000 BioRS Dual System. The Thermo Scientific Acclaim RSLC 120 C18, 2.2 µm, 2.1x250 mm column was operated at 80 °C, to obtain high resolution separation of the complex mixture. Due to the high temperature in the column, the pre-heating of the mobile phase was essential to avoid thermal mismatch between the mobile and the column. A pre-column heater 2 µL biocompatiblewas used to achieve thermal equilibration between column and mobile phase.
