For years, mass spectrometry analysis of intact proteins has suffered from important technical limitations that, on the other hand, were only marginally impacting peptide-centric proteomic approaches. Recently, this performance gap has been partially filled by the introduction of high-resolution mass analyzers, a variety of new ion activation techniques, and novel signal processing algorithms. Nonetheless, some of the old problems affecting top-down proteomics cannot be solved simply by applying these improved technological resources to standard data acquisition schemes. Therefore, to circumvent some of these issues – specifically those linked to the characterization of large proteoforms – we will describe potential solutions using alternative strategies based on ion manipulation, gas-phase fractionation and targeted data acquisition.
Prof. Luca Fornelli
Luca Fornelli earned a B.Sc. and M.Sc in Biotechnology at the University of Padova in Italy. He then moved to the École Polytechnique Fédérale de Lausanne (EPFL), Switzerland, to obtain his Ph.D. in Chemistry under the supervision of Dr. Yury Tsybin, where he worked on the analysis of intact antibodies using Fourier transform mass spectrometry. In 2014 he obtained a postdoctoral mobility fellowship from the Swiss National Science Foundation and joined the group of Dr. Neil Kelleher at Northwestern University. He joined the University of Oklahoma in 2019, where is currently an Assistant Professor at the Departments of Biology and Chemistry & Biochemistry. Luca’s research is focused on the qualitative and quantitative characterization of large intact proteoforms through high-resolution mass spectrometry, using a variety of tools – including novel ion dissociation methods, gas-phase fractionation and ion-ion reactions –, to develop new applications of top-down proteomics for advancing basic and translational sciences. Read more about his work at https://www.ou.edu/cas/biology/people/faculty/luca-fornelli