Development of MRM Methods for Monoclonal Antibodies Using Skyline

Introduction

Monoclonal antibodies, or mAbs, have been used for over a decade in the treatment of a number of diseases but predominantly in the treatment of cancer and autoimmune diseases. Quantifying therapeutic mAbs in biological samples has been traditionally addressed by ligand binding assays (LBA’s), however, there are major limitation in terms of extended method development times, reagent procurement, and matrix effects. LC-MS/MS methods are emerging as an alternative approach to LBA’s and this paper describes the application of MRM methods to the quantitation of peptide fragments containing β-amyloid antibody (6E10) CDR’s (complementarity determining regions). CDR’s are targeted by LC-MS/MS as antibodies share a conserved sequence and are only differentiated by the three loops (CDR1-3) which are diverse and contain antibody specific peptides. By selectivity detecting CDR1-3 by LC-MS/MS helps to characterize and quantify disease specific antibodies isolated from patient sera or plasma and can be used to identify differing therapeutic agents.

April 1, 2016 GMT