Please join us on Tuesday, February 12th, 2019 at Northwestern University, Ryan Hall 4003. Dinner starts at 6:30 PM with the seminar immediately following at 7:00 PM. Please be sure to register so our sponsors know how much food to order! We hope you can join us! To register click here
Speaker: Luca Fornelli, Assistant Professor at the Department of Biology, the University of Oklahoma (Norman, OK)
Title: Proteoform cations and their new, old friends: UV photons and perfluorinated anions
Abstract: Top-down mass spectrometry (TDMS) is a powerful tool to characterize intact proteoforms. To be routinely applied as a high-throughput proteomic technique, TDMS has to face two major challenges: (i) obtaining high (possibly complete) sequence coverage of the proteoforms selected for tandem mass spectrometry and (ii) becoming capable of easily analyzing mixtures of proteoforms > 30 kDa (the current mass limit). As it often happens, solutions to new problems (might) come from the past. Examples of ultraviolet photodissociation (UVPD) of polypeptide cations were initially reported in the eighties, but this ion fragmentation technology has been rediscovered and applied to high-throughput proteomics only recently, particularly using 193 nm photons. Similarly, ion-ion proton transfer reactions (PTR) have been described by Stephenson and McLuckey in the nineties, but have found limited application in the “omics” sciences until now. Here, we will discuss the latest advances in the application of UVPD to top-down proteomics using a 213 nm laser, with special focus on UVPD-generated product ions & their relative abundances, to determine how to properly analyze UVPD spectra in proteomic experiments. Furthermore, we will illustrate the first results of a novel application of PTR to the characterization of large proteoforms (up to 60 kDa) from Pseudomonas aeruginosa.
Bio summary: Luca Fornelli obtained his PhD in Chemistry and Chemical Engineering from the Ecole Polytechnique Federale de Lausanne (EPFL), Switzerland, in 2014. Under the supervision of Dr Yury Tsybin, he developed new strategies for middle-down and top-down characterization of therapeutic antibodies. As a postdoc in the laboratory of Dr Neil Kelleher at Northwestern University he then applied the top-down approach to both high-throughput and targeted proteomics.