Presented by: Gavin Reid, Associate Professor at Michigan State University
A large number of studies have demonstrated that disruption of lipid metabolism or signaling pathways can play a key role in the onset and progression of human disease, including cancer and diabetes. Thus, a comparative analysis of changes in individual lipids or lipid profiles (i.e., the lipidome) between normal and diseased cells, tissues, organs, or accessible bodily fluids (e.g., tumor interstitial fluid, blood plasma or serum), may enable the identification and characterization of lipids that can serve as effective biomarker signatures of the disease. In this presentation, the development and application of a straightforward and high throughout analysis strategy consisting of high-resolution ‘shotgun’ mass spectrometry (MS), ‘targeted’ tandem mass spectrometry (MS/MS), functional group specific chemical modification and in situ liquid extraction of cell culture samples is described for the comprehensive identification, characterization and quantification of multiple lipid classes from within a colon adenocarcinoma cell line, SW480, and its metastasized derivative, SW620.
Presented by: Ljiljana Paša-Tolić, Ph.D., Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory
Description: Obtaining extended sequence coverage of intact proteins and characterizing multiple post-translational modification (PTM) sites can pose key technical challenges. In this webinar, we describe how we have implemented Advion’s TriVersa NanoMate® and RePlay® technologies to address these issues using an integrated top-down/bottom-up approach.
We demonstrate how the TriVersa NanoMate’s capabilities of chip-based electrospray ionization (ESI) and fraction collection are used to profile isoforms, further investigate targeted proteins at the intact level using ECD or CID, perform bottom-up proteomics on a collected fraction to get confident protein identifications, perform targeted MS/MS on particular fractions of interest, and better characterize where PTMs may have occurred.
We also discuss how RePlay simplifies the data analysis process by allowing you to see the correlation between the intact protein and the observed peptides, even when you are sample limited. Using RePlay, we incorporated an on-line digestion strategy with the integrated top-down/bottom-up approach. This technique allows you to implement LC-MS/MS during both the intact run and a secondary RePlay run with digestion, so you can see the correlation in time between the observed parent ions and the digested ions as well as the MS/MS of the fragment ions.
Martin NJ, Bunch J, Cooper HJ J Am Soc Mass Spectrom. 2013 Aug;24(8):1242-9
M. Wisztorski; B. Fatou; J. Franck; A. Desmons; I. Farre; E. Leblanc; I. Fournier; M. Salzet Proteomics Clin. Appl. 2013, 7, 1-7
V. Kertesz; G.J. Van Berkel Bioanalysis 2013 Apr;5(7):819-26. doi: 10.4155/bio.13.42.
Ellis, S.R.; Ferris, C.J.; Gilmore, K.J.; Mitchell, T.W.; Blanksby, S.J.; In Het Panhuis, M.
Anal Chem. 2012 Nov 7. [Epub ahead of print]
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Anal Chem. 2012 Oct 30. [Epub ahead of print]
Cubrilovic, D.; Biela, A.; Sielaff, F.; Steinmetzer, T.; Klebe, G.; Zenobi, R.
J Am Soc Mass Spectrom. 2012 Oct;23(10):1768-77. Epub 2012 Aug 7
Edwards, R.L.; Griffiths, P.; Bunch, J.; Cooper, H.J.
J Am Soc Mass Spectrom. 2012 Nov;23(11):1921-30. doi: 10.1007/s13361-012-0477-9. Epub 2012 Sep 20.
Annalisa Arcella, Guillem Portella, Maria Luz Ruiz, Ramon Eritja, Marta Vilaseca, Valérie Gabelica, and Modesto Orozco
Journal of the American Chemical Society 2012 134 (15), 6596-6606
