Around for more than a century the analytical technique of mass spectrometry is blooming more than ever, and applied in nearly all aspects of the natural and life sciences. In the last two decades mass spectrometry has become routine for the high-throughput analysis of peptides and glycans, and to a lesser extent glycopeptides. However, also intact proteins and even complete protein complexes can nowadays be analyzed. In this lecture, I will describe the emerging role of mass spectrometry with its different technical facets in molecular and structural biology, focusing especially on the analysis of intact glycoproteins. Moreover, I will describe how we use native mass spectrometry to study dynamic protein assemblies, for instnace those involved in complement activation.
Recent developments in mass spectrometry technology have allowed us to analyze intact native glycoproteins and protein complexes using Q-ToF and Orbitrap mass analyzers with very high sensitivity and mass resolving power, enabling us to profile the quality and biosimilarity of protein biotherapeutics, in their native state without requiring much sample preparation. In detail, I will demonstrate how native mass spectrometry can be combined with middle-down proteomics to profile complex structures of various glycoproteins, focusing on mAbs, Erythropoietin and plasma proteins. Thereby I will address the question of how unique each person is, as viewed from each individual’s glycoproteome.