This application provides a fast and accurate way for biopharmaceutical chemists to monitor PEGylation reactions by using HPLC with complementary UV/Vis and charged aerosol detection.Separation on the MAbPac RP resolves PEGylation reagents, unreacted IgG, and PEGylated IgG products that elute as a broad hump.

Size exclusion chromatography (SEC) is one of the major analyses in biotherapeutics characterization. Here we show a MAbPac SEC-1 column test with the Thermo Scientific Vanquish Flex UHPLC system.

Hydrophobic interaction liquid chromatography (HIC) is one of the major analyses in biotherapeutics characterization. Here we show a high resolution separation of proteins and a mAb in 20 minutes with the Thermo Scientific Vanquish Flex UHPLC system. The unique chemistry of the MAbPac HIC-20 column allows the analysis of hydrophilic proteins such as myoblogin and ribonuclease A as well as more hydrophobic mAbs.

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.

Glycans released from bovine fetuin by PNGase-F are separated by gradient elution on the Thermo Scientific Dionex GlycanPac AXR-1 column and detected by charged aerosol detection on the Corona Veo RS.