Through advances in technologies the role of mass spectrometry in structural and molecular biology is rapidly expanding. Especially hybrid MS approaches combining top-down, middle-down and bottom-up proteomics analysis with native MS, HD exchange MS or XL-MS assist us to provide unprecedented detail on the structural and function heterogeneity present in dynamically evolving proteins and protein assemblies, including complex plasma glycoproteins but also in intact ribosomes and viruses.
In this talk I will describe novel developments in MS instrumentation for native MS and top-down MS, especially new modifications to an Orbitrap based instrument that offers high-sensitivity and mass resolving power, allowing an in-depth detailed analysis of highly glycosylated plasma glycoproteins, protein assemblies up to even whole intact ribosomes and viruses.
I will describe how we use these new mass analysers to characterize in depth pharmaceutical proteins and plasma glycoproteins. Thereby we focus on how the proteoform profiles of targeted plasma glycoproteins respond to perturbations such as vaccination, sepsis and cancer.
I will also describe the latest developments in the lab around cross-linking mass spectrometry where we attempt to perform proteome-wide cross-linking using XlinkX, extending depth by imporved fragmentation schemes and our in-house developed enrichable cross-linker PhoX.
Finally, I will highlight some of the latest developments in using hybrid fragmentation methods including HCD, ECD/ETD and UV/IRMPD laser photodissociation for native and top-down proteomics.