Presented by: John P. Shockcor, Director of Life Sciences Business Development, Waters Corp., Visiting Fellow, Dept. of Biochemistry, University of Cambridge, UK
Description: Profiling low level components in a complex mixture of small molecules can be a challenging task. Although it may be possible to detect many low level components in a complex mixture, characterization is often hindered because fragmentation of these low level components yields peaks below the limit of detection. This problem can be alleviated by using a TriVersa NanoMate assisted approach. In this webinar we will describe how a TriVersa NanoMate coupled to a SYNAPT G2 Hybrid QTof Ion-Mobility Mass Spectrometer can provide critical fragmentation information needed to characterize low level components in lipidomics, drug metabolism studies and natural product profiling. This approach is ideally suited to the use of time-aligned-parallel fragmentation (TAP) which are illustrated by a number of examples.
Presented by: Dr. Daniel Eikel, Sr. Application Scientist – Advion, Inc.
Description: Liquid extraction surface analysis mass spectrometry (LESA-MS) is a novel mass spectrometry-based surface-profiling technique (Kertez, et al., 2009) that can be utilized in drug distribution and metabolism studies. Potential advantages of LESA-MS are as follows:
- No radiolabeled compound is required
- Its overall sensitivity appears favorable compared to autoradiography or matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI)
- No additional sample preparation such as MALDI matrix application is required.
In this webinar, Dr. Daniel Eikel expands upon his publication (Henion, et al., 2011), discussing how LESA-MS can provide complementary information to the gold standards in this field: quantitative whole body autoradiography (QWBA) and whole organ LC-MS/MS, or other MS based approaches such as MALDI-MSI. He demonstrates how they have evaluated LESA-MS by studying the drug distribution and metabolism of terfenadine in mouse, and through direct comparison, proved that LESA-MS appears to be more informative than a comparable MALDI-MSI experiment and reflects the literature-known metabolism and distribution pattern for terfendine and its metabolite fexofenadine.
Presented by: Thomas Covey, Principal Research Scientist at AB SCIEX
Thomas Covey describes how various organizations are incorporating the fast, simple, and direct method of LESA – Liquid Extraction Surface Analysis – into their labs. He shows data that illustrates how you can see more analytes with better sensitivity by combining LESA with mass spectrometry and chip-based nanoESI via Advion’s TriVersa NanoMate. This new analytical technique has a wide range of applications for sample analysis from a variety of surfaces e.g. tissue slices, TLC plates, MALDI plates, and Dried Blood Spots.
Dr. Covey also demonstrates the latest LESA upgrade – LESA Points software that provides simple, point-and-click surface analysis. Users can scan a digital image of their samples and select sampling positions within 90-μm resolution. The new upgrade also provides a specially-developed mounting block that keeps all kinds of surfaces at a fixed height for simple method set-up.
In this webinar, Dr. Han describes how he uses the TriVersa NanoMate’s chip-based nanoelectrospray ionization capabilities in infusion mode to obtain more information from complex samples and faster lipids analysis with no sample-to-sample carryover. The interests of Dr. Han’s laboratory have been focused on the altered lipid metabolism, trafficking, and homeostasis under patho(physio)logical conditions. Currently, there are three specific areas explored in his laboratory including (1) extension of the shotgun lipidomics technology for increased penetrance into the low abundance regime of a cellular lipidome with emphasis on high throughput, and bioinformatics; (2) investigation of the biochemical mechanisms underlying the altered lipid content and composition in metabolic syndrome; and (3) identification of the biochemical mechanisms responsible for the sulfatide depletion and ceramide elevation at the very earliest stages of Alzheimer’s disease.
Presented by: Xianlin Han, Ph.D., Professor, Diabetes and Obesity Research Center, Sanford-Burnham Medical Research Institute
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]
