Fast Online Desalting of mAbs Using a Reversed-Phase Desalting Cartridge for LC-MS Analysis

Instrument Type: LCMS

This collateral describes a LCMS method for high throughput intact antibody characterization, capable of delivering the intact mass and the relative glycoform abundance of a mAb within 5 minutes.The desalting method optimization was conducted on a 2.1 × 10 mm Thermo Scientific™ MSPac™ DS-10 cartridge using conventional water/acetonitrile-based separations on the Thermo Scientific™ Vanquish™ UHPLC system and MS detection using the Thermo Scientific™ Q Exactive™ Plus Hybrid Quadrupole-Orbitrap™ mass spectrometer.

Reversed-Phase Separation of Intact Therapeutic Antibodies Using the Vanquish Flex UHPLC System

Instrument Type: HPLC (Biocompatible)

Reversed-Phase chromatography (RP) of intact antibodies is one of the major analyses in biotherapeutics characterization. Here we show RP of several intact antibodies with The Thermo Scientific Vanquish Flex UHPLC in combination with the Thermo Scientific MAbPac-RP column. Column temperature was 80 °C; the mobile phase was pre-heated at the same temperature. Four antibodies were eluted with 10 minutes gradient. The elution of the intact antibodies resulted in very sharp and symmetrical peaks. Separation with the MAbPAc-RP were able to distinguish between a reference and a stressed antibody.

Improved analysis of intact proteins using a Thermo Scientific Accucore 150-C4 150 Å pore diameter nanoLC column

Instrument Type: HPLC

The Thermo Scientific HPLC system is applied for the analysis of intact proteins (*indicates an impurity from carbonic anhydrase). The separation is performed on a Thermo Scientific Accucore 150-C4 150 Å pore diameter nanoLC column, with UV detection at 214 nm. The 150 Å pore size enables the effective analysis of molecules unable to penetrate into smaller diameter pores, whilst the low hydrophobicity C4 phase results in protein separation by hydrophobicity.

Improved analysis of intact proteins using a Thermo Scientific Accucore 150-C4 150 Å pore diameter HPLC column

Instrument Type: HPLC

The Thermo Scientific HPLC system is applied for the analysis of intact proteins. The separation is performed on a Thermo Scientific Accucore 150-C4 150 Å pore diameter HPLC column in under 10 minutes, with UV detection at 214 nm. The 150 Å pore size enables the effective analysis of molecules unable to penetrate into smaller diameter pores, whilst the low hydrophobicity C4 phase results in protein separation by hydrophobicity.