Triple quadrupole mass spectrometer is the LC/MS platform of choice in bioanalysis of peptide or protein therapeutics, in which quantitation is usually accomplished by MRM detection of intact peptide or signature peptides of a targeted protein via enzymatic digestion. In some cases however peptides do not generate useful fragment ion(s) allowing for sensitive or selective MRM analysis. HRMS-based accurate analysis has been recently employed to solve this problem. In this presentation, we describe an alternative to HRMS by combining differential mobility spectrometry (DMS) and triple quadrupole-based multiple ion monitoring (MIM) for quantitation of peptides that cannot be effectively analyzed by MRM. Preliminary results suggest that the DMS-MIM workflow had assay LLOQ sensitivity and selectivity similar to HRMS results. This strategy enabled triple quadrupole MS instruments to perform the bioanalysis of poorly fragmented peptides. In addition, a workflow using DMS to reduce interference in peptide analysis by MRM is presented. Such orthogonal methodologies would greatly benefit bioanalytical labs in several aspects, including cost saving, method development and validation, and GLP compliance.
What will you learn?
Bioanaytical quantitation of proteins
Orthogonal options in bioanalytical quantitation
Comparison of HRMS and QTRAP
Who should attend?
Laboratory Managers/Group Leaders/Sr Scientists
Bioanalytical Laboratory Directors
January 27, 2015 7:00 a.m. PST 10:00 a.m. EST 15:00 GMT