CHARACTERIZATION OF A MONOCLONAL ANTIBODY (mAb) USING MULTIPLE FRAGMENTATION TECHNIQUES AND NOVEL FOURIER TRANSFORM DATA PROCESSING SOFTWARE

摘要

Antibodies represent a class of proteins that are the key to immunological defense systems. They bind to an antigen (protein, glycoprotein, DNA, etc.) with a high degree of specificity and neutralize foreign objects, making them extremely valuable for use in diagnostics, general research, and for therapeutics. Since their introduction in the late 1980s, monoclonal antibodies (mAb) have become popular therapeutic candidates due to high specificity and low side effects. Most of the clinically approved mAbs, IgG1s are commonly to be heterogeneous glycosylated which are not efficiently identified in proteoforms by conventional bottom-up analysis. Recently, top- and middle-down analyses have been shown as efficient tools for identification of PTMs that are often lost when proteins are analyzed after proteolysis. We employed intact protein analyses together with multiple fragmentation techniques, electron transfer dissociation (ETD) and MALDI-in-source decay (ISD), by use of FT-ICR and/or Orbitrap to provide improved sequence coverage and comprehensive glycan profiling of two large proteins, IgG1 and IgG1-fusion proteins.