Analysis of Monoclonal Antibody (mAb) and Related Biologics by LC and LC-MS Platforms

Overview

Title: Analysis of Monoclonal Antibody (mAb) and Related Biologics by LC and LC-MS Platforms 

Duration: 21 minutes

Available On Demand

Summary

Monoclonal antibodies (mAbs) are a class of biotherapeutics that are rapidly gaining traction. The higher-order structures of mAbs play a crucial role in determining their efficacy and safety. It is crucial that mAbs are accurately characterized before they can become biotherapeutics. A better understanding of mAbs helps craft better bioprocesses, formulas, and dosages. In this study, a recombinant human IgG NIST mAb reference standard was analyzed using high resolution quadrupole time of flight liquid chromatography-mass spectrometry (QTOF-LCMS). An innovative proteolysis method called nano-surface and molecular-orientation limited proteolysis (nSMOL) selectively cleaves Fab regions of mAbs. Regardless of the type of antibodies scientists use, nSMOL can be used to develop analytical methodologies. Triple quadrupole liquid chromatography-mass spectrometry (TQ-LCMS) permits multiple reaction monitoring (MRM) to determine the level of fragmentation in peptides derived from Fab. Furthermore, a protocol for selecting pharmacokinetically suitable signature peptides was developed using nSMOL and TQ-LCMS.