This study shows a fully automated UHPLC setup applying a simple and efficient online solid phase extraction method. Showcase assay performance by using a column cartridge for sample trapping and a Thermo Scientific™ MAbPac™ RP column for separation with water/acetonitrile-based gradients. Leverage the new technologies such as the Thermo Scientific™ Vanquish™ UHPLC platform in combination with the Thermo Scientific™ Q Exactive™ HF Hybrid Quadrupole Orbitrap mass spectrometer.

The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Ion Exchange analysis of Rituximab. The separation was performed on a 3 micron Thermo Scientific MabPac SCX-10 column with a UV detection at 280 nm. With a run time of 16 minutes, the method allows easy separation of acidic and basic charge variants of Rituximab

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.

The Thermo Scientific Ultimate 3000 BioRS Dual system is used in combination with the Thermo Scientific MabPac SCX-10 RS 2.1x50mm to run fast pH-gradient ion exchange of cetuximab. The pH gradient is generated by using the CX-1 pH-gradient buffers.A fast generic screening method is run with a 10 minutes gradient from pH 5,6 to pH 10,2. The generic screening method isoptimized to achieve high throughput, high resolution separation of the charge variants.

The Thermo Scientific Vanquish UHPLC system is used in combination with the Thermo Scientific MabPac SCX-10 RS 2.1x50mm to run fast pH-gradient ion exchange of four monoclonal antibodies. The pH gradient is generated by using the CX-1 pH-gradient buffers. A fast generic screening method is run with a 10 minutes gradient from pH 5,6 to pH 10,2. The generic screening method was afterward optimized for each antibody to achieve high throughput, high resolution separation of the charge variants.