Tutorial & eScience Café Seminars

At-A-Glance

MONDAY, JULY 12

  • Tutorial on “Capillary Iso-Electric Focusing (CIEF) – Prime Methodology for Protein Characterization” (presented by Gerard Rozing)
  • eScience Café Seminar on “Combining the Power of a Core-Shell Particles and Advanced Stationary Phase Selectivity to Improve Micro and Nano Flow Separations" (sponsored by Phenomenex, Silver sponsor)

TUESDAY, JULY 13

  • eScience Café Seminar on "Microscale Separations at Nanoscale" (sponsored by VICI, Gold sponsor)
  • eScience Café Seminar on "Is Structures for Lossless Ion Manipulation (SLIM) a One Trick Pony or a One Stop Shop for Ion Mobility-Mass Spectrometry Analysis?" (sponsored by MOBILion Systems, Inc., Silver sponsor)

WEDNESDAY, JULY 14

  • eScience Café Seminar on “Latest Applications of 4D-Proteomics using Trapped Ion Mobility on the timsTOF Pro 2” and 4D Proteomics – Dissecting the 3D Structure of Proteins through Ion Mobility Enhanced Crosslinking Mass Spectrometry” (sponsored by Bruker, Gold sponsor)
  • eScience Café Seminar on “Comprehensive, 15-min Charge Variant Analysis of Biotherapeutics with a Microfluidic Chip-Based Integrated iCIEF-MS System” (sponsored by SCIEX, Gold sponsor)

THURSDAY, JULY 15

  • eScience Café Seminar on “Advancing Denaturing and Native Top-down Proteomics Analysis using CE-MS” (sponsored by Agilent, Silver sponsor)

 

Monday, July 12 at 12:10 pm – 12:55 pm

Free Tutorial

Capillary Iso-Electric Focusing (CIEF) – Prime Methodology for Protein Characterization

Presented by Dr. Gerard Rozing, ROZING.COM Consulting, Karlsruhe, GERMANY

Since the inception of iso-electric focusing (IEF) as a high-resolution electrophoretic technique for the separation of amphoteric substances by Tiselius and Vestenberg, it has become an indispensable method for the separation of protein mixtures. Initially, IEF was executed on a flat surface coated with a gel mixed with ampholytes to establish the pH gradient. The sample is applied to the center of the plate. This so-called slab gel IEF became more sophisticated by using flat surfaces with an immobilized pH gradient (IPG). With the introduction of instruments for Capillary Electrophoresis, the technique was adapted to allow execution in-the capillary and subsequent, online optical detection (capillary iso-electric focusing, CIEF). This was followed by dedicated whole-column-image-detection electrophoresis systems (iCIEF). With the commercialization of CIEF, the methodology has evolved into a recipe-driven, black-box approach. The users strictly follow a protocol provided by the system supplier. Therefore, the basic understanding of the separation mechanism on IEF has faded. Diagnosis and troubleshooting when “it does not work” become an insurmountable hurdle, and the costly help of the system supplier is called. Therefore, there is a continuing need and effort for teaching the basic principles of iso-electric focusing, especially when applied in capillaries (CIEF). In this tutorial lecture, the author will emphasize on: (a) Basics of iso-electric focusing and in capillary execution. (b) Method development of CIEF. (c) Instrumentation for CIEF and iCIEF. (d) Tools for capillary iso-electric focusing method development. (e) Preparative CIEF. (f)  Coupling of CIEF with Mass Spectrometric Detection. In the spirit of the MSB symposia series, the author will fill 2/3 of the allocated time on presentation leaving 1/3 for Q&A.

Rozing

Gerard Rozing. 1977: Ph.D. Amsterdam University. Organic Chemistry and Chemical Engineering. 1977-1979: Post-doctoral research University of Ghent, Belgium and Amsterdam University. 1979-1999: Hewlett-Packard, Waldbronn, Germany. R&D Chemist, Group & Project Leader, HPLC columns, HPLC system, CE capillaries and capillary electrophoresis system development. 2000-2012: Agilent Technologies, University Relations and External Scientific Collaborations Manager, Agilent Research Fellow. 2012: Owner, ROZING.COM Consulting. Member of the Strategy Advisory Board of PharmaFluidics, Ghent, Belgium, Scientific Consultant with Advanced Electrophoresis Solutions, Cambridge, Canada. Co-organizer, HPLC, MSB, ITP and ISC Symposium series. Website https://www.rozing.com

Free eScience Café Seminars
Monday, Tuesday, Wednesday, Thursday

Monday, July 12 at 12:10 pm – 12:55 pm

Free eScience Café Seminar sponsored by PHENOMENEX

Combining the Power of a Core-Shell Particles and Advanced Stationary Phase Selectivity to Improve Micro and Nano Flow Separations

Presented by Jason A. Anspach, Global Product Manager HPLC, Phenomenex, Torrance, CA, USA

Phenomenex logo

It has long been shown that by improving separation quality one can substantially improve the quality of the mass spectrometry data. Improvements including the ability to resolve and detect isobaric compounds, reducing spectrum complexity for the improved sampling of unknowns, or improvements in detection limits can all be accomplished via improvements in LC separations. The two most powerful tools to improve LC separations are improvements in column efficiency, and enhancements in stationary phase selectivity. While there are a vast number of publications discussing methods to improve both efficiency and selectivity in analytical scale separations, these topics have largely been ignored in micro and nano scale separations. In this workshop we will show how the use of core-shell based HPLC materials, long known to significantly improve efficiency in analytical HPLC, and alternate stationary phase bonding chemistries can be used in micro and nano scale LC/MS analysis to improve separation quality and provide better sensitivities.

Anspach

Jason Anspach received his BS in Chemistry from Binghamton University, The State University of New York and his Ph.D. from The State University of New York at Buffalo where he worked under the advisement of Luis A. Colón. His work there centered on improving column performance through novel column packing methods and the use of Sub 2 µm packing materials. He has continued that work since joining Phenomenex in 2005 leading an R&D team advancing column packing and hardware technologies. In 2019 he joined the product management team in the role of Global Product manager for HPLC.

Tuesday, July 13 at 12:35 pm – 1:20 pm

Free eScience Café Seminar sponsored by VICI

Microscale Separations at Nanoscale

Presented by Jennifer Copeland, LC Product Manager, VICI, Houston, TX, USA

Gold Sponsor - VICI

While microscale separations have been an intense area of study over the past few decades, application of these techniques for single-cell analysis of proteomes is newly possible. Novel efforts into cellular study are ongoing and wide-ranging, and there have been numerous methods of looking at protein makeup of an individual cell. Each of these technologies requires the ability to move nearly all (if not all) of a tiny sample to a separation modality and then to a detector. Moving to nanoscale has been challenging in multiple aspects, and this workshop will go over some of the history and methods developed, while discussing technology involved in ultra-high resolution packed-bed capillary chromatography. VICI’s True NanoTM product line solves many common problems facing chromatography while taking advantage of column and detector improvements over the past few years. Join us as we discuss bringing the analytical power of liquid chromatography into the single-cell present.

Copeland_SpeakerPhoto

Jennifer Copeland has been working in analytical separations and instrumentation for nearly 20 years, with experience in pressure and current-driven separations, mass spectroscopy, NMR, direct and indirect light detection, and data analysis. Dr. Copeland has also worked on platform development for protein-ligand binding screens including compound library development, protein binding-site comparisons, single cell RT-PCR profiling, extra-cellular protein detection and isolation, and novel nano-scale UHPLC systems. She received her Ph.D. in Chemistry from the University of Nebraska in Lincoln in 2013, and has been working at VICI for nearly seven years. As the Liquid Chromatography Product Manager, Dr. Copeland works as the liaison between laboratory scientists and VICI's engineering teams to facilitate product testing and development.

Tuesday, July 13 at 12:35 pm – 1:20 pm

Free eScience Café Seminar sponsored MOBILion Systems, Inc.

Is Structures for Lossless Ion Manipulation (SLIM) a One Trick Pony or a One Stop Shop for Ion Mobility-Mass Spectrometry Analysis? 

Presented by Daniel DeBord, Director of R&D, MOBILion Systems, Inc., Chadds Ford, PA, USA

MOBILion_Color_Horizontal_Logo

Since 2014 SLIM has demonstrated unprecedented levels of ion mobility resolution and the potential impact of this resolution on both the depth and throughput of analysis. Integration has focused on the combination of high-resolution ion mobility (HRIM) with high resolution mass spectrometry (HRMS) in the form of time-of-flight instrumentation. This system configuration provides broad spectrum acquisition with fast analysis times, making it most suitable for discovery or characterization analyses where the goal is to see “everything” with as much specificity as possible. MOBILion has been working to evolve SLIM functionality to support targeted applications where the goal is to focus on certain analytes, and to be able to detect them as accurately and sensitively as possible, even within complex biological matrices. This presentation provides an overview of various designs of SLIM and shows how this platform technology can be tailored to enhance performance of all types of mass spectrometric detection. 

Daniel DeBord

Daniel DeBord is the Director of R&D for MOBILion Systems where he helps guide the development and commercialization of SLIM- based analytical instrumentation. Since receiving his Ph.D. in Analytical Chemistry from Texas A&M University, he has served as the Associate Director of the Advanced Mass Spectrometry Facility at Florida International University and held industrial positions at BASF and 1st Detect Corporation, working on a variety of mass spectrometry related applications and instrumentation projects. His extensive experience in the ion mobility and mass spectrometry fields includes designing miniaturized MS-based detection systems for non-traditional markets as well as contributions to early Trapped Ion Mobility Spectrometry experiments.

Wednesday, July 14 at 12:35 pm – 1:20 pm

Free eScience Café Seminar sponsored by BRUKER

Latest Applications of 4D-Proteomics using Trapped Ion Mobility on the timsTOF Pro 2

Presented by Gary Kruppa, Vice President Proteomics at Bruker Daltonics Inc., and Managing Director of Bruker S.R.O., Billerica, MA, USA

 

4D Proteomics – Dissecting the 3D Structure of Proteins through Ion Mobility Enhanced Crosslinking Mass Spectrometry

Presented by Richard Schletema, Assistant Professor, Pharmaceutical Sciences, University of Utrecht, Utrecht, THE NETHERLANDS

MSB-Conferences.org

Gary Kruppa, Ph.D. will present an update on the newly introduced timsTOF Pro 2 and the timsTOF SCP. The use of trapped ion mobility separation (TIMS) to enable Parallel Accumulation Serial Fragmentation for DDA PASEF will be explained. Then the principles and performance of the dia-PASEF and prm-PASEF methods will be covered. Updates on the latest exciting performance for ultra-high sensitivity and single cell proteomics will be shown. In the second half, Professor Richard Scheltema of the University of Utrecht will present results on ion mobility enhanced crosslinking mass spectrometry (caps-PASEF) to study the 3D structure of proteins. The use of the PhoX crosslinker which contains a phosphonate moiety that allows enrichment of species that react with the crosslinker will be introduced. The use of the caps-PASEF method to further focus on crosslinked species will be shown and applications to the study of proteins 3D protein structures will be discussed.    

Kruppa
scheltema

Gary Kruppa serves as the Vice President for proteomics at Bruker Daltonics Inc. He manages a growing global team of applications, field applications and applications development scientists, focused on developing new proteomics applications for the timsTOF Pro, and supporting customers and collaborators. Dr. Kruppa earned his bachelor’s degree in chemistry at the University of Delaware and his Ph.D. in chemical physics at CalTech. Dr. Kruppa has been working with Bruker since 1991, where he first worked on the FTMS product line, later serving as Vice President for FTMS. From 2001 to 2004, Dr. Kruppa ran his own research lab at Sandia National Labs, where he and his team developed novel methods and reagents for chemical crosslinking of proteins for the study of protein structure. In 2004 he rejoined Bruker, where he served as Vice President for Business Development until 2014. From 2014 to 2016 Dr. Kruppa worked as CEO of MRM Proteomics Inc., to commercialize kits for quantitative MRM proteomics analysis of potential biomarkers. Dr. Kruppa returned to Bruker in 2016 to serve as Vice President for proteomics, where he helped launch the revolutionary timsTOF Pro QTOF Mass Spectrometer powered by the PASEF method.

Richard Scheltema currently runs his own, independent research group at Utrecht University. After finalizing his Postdoc working on LC-MS/MS platforms within the group of Prof. Dr. Matthias Mann (Max Planck Institute of Biochemistry, Germany), he started the Scheltema laboratory at Utrecht University. The focus lies on structural proteomics applied to medium and high complexity protein environments like the ECM and membrane proteins. Mass spectrometry-based approaches are employed that are independent of protein size, deal with high complexity mixtures and play well with other structural biology techniques like Cryo-EM and crystallography to uncover novel and exciting biology. The technological developments and their applications have been published in journals like Nature ProtocolsNature Communications, Molecular Cell, Cell ReportsACS Central Science, and PNAS and additionally have been commercialized and are worldwide in use.

Wednesday, July 14 at 12:35 pm – 1:20 pm

Free eScience Café Seminar sponsored by SCIEX

Comprehensive, 15-min Charge Variant Analysis of Biotherapeutics with a Microfluidic Chip-Based Integrated iCIEF-MS System

Presented by Maggie A. Ostrowski, Intabio, now part of SCIEX, Fremont, CA, USA

sciex logo updated with tagline_CMYK color

Charge heterogeneity analysis is essential for the successful development and production of therapeutic monoclonal antibodies (mAbs) in the biopharmaceutical industry. Charge heterogeneity of mAbs can result from cellular processes, chemical degradation, and production conditions during manufacturing. These changes can impact the therapeutic mAb’s efficacy, safety and potency. As a result, the complex microheterogeneity of mAbs requires in-depth structural characterization for critical quality attribute (CQA) assessment. Capillary isoelectric focusing (cIEF) is a technique for separating charge variants where mAb modifications and degradation products affect the mAb’s isoelectric point. cIEF has been widely adopted within the biopharmaceutical industry. However, when a change in a mAb is measured by cIEF, the process of identifying the post-translation modification that resulted in the change can be time-consuming and laborious, often requiring separation and fraction collection, followed by offline analysis by mass spectrometry. The integration of cIEF with MS results in a powerful analytical approach for more comprehensive and rapid mAb characterization. This presentation will summarize the development of our imaged cIEF-MS system designed specifically to enable full mass characterization of iCIEF peaks in a single 15-minute assay. The iCIEF separation, chemical mobilization and electrospray ionization (ESI) functions are integrated into a microfluidic chip, enabling automation of the workflow and direct interfacing with high-resolution QTOF systems for mass determinations and charge variant characterization. Several examples of charge variant characterization demonstrating high-resolution cIEF separation coupled with sensitive detection and mass identification of major and minor isoforms will be highlighted.

Ostrowski

Maggie A. Ostrowski is the Senior Marketing Manager at SCIEX in the CE and Biopharma Business Unit. Before the acquisition by SCIEX, Maggie was responsible for strategic marketing for the iCIEF-MS system at Intabio, Inc. She is passionate about developing customer-focused mass spectrometry solutions and enabling technologies for the characterization of biotherapeutics. Prior to her role at Intabio, Maggie held various roles in mass spectrometry product management and R&D at Agilent Technologies focused on the development and marketing of analytical technologies with an emphasis on microfluidics-based bioanalytical measurement and cell analysis. She has authored numerous scientist publications and holds several patents. She earned a PhD in Chemical Engineering from Stanford University and a BS in Bioengineering from UC Davis. 

Thursday, July 15 at 12:35 pm – 1:20 pm 

Free eScience Café Seminar sponsored by AGILENT

Advancing Denaturing and Native Top-down Proteomics Analysis using CE-MS

Presented by Liangliang Sun, Assistant Professor, Department of Chemistry, Michigan State University, East Lansing, MI, USA

Agilent_Logo_RGB

Studying the function of proteins in cells is vital for understanding the underlying molecular mechanisms of disease and development. Posttranslational modifications (PTMs) can influence protein conformations and function. Understanding protein function requires a deeper characterization of proteoforms and their associated protein complexes. Top-down proteomics (TDP) aims to delineate proteoforms and protein complexes on a global scale and in discovery mode. It has achieved substantial progress in the last decade. However, many challenges remain, including but not limited to high-capacity separation and extensive fragmentation of proteoforms and protein complexes. In this talk, we will introduce capillary electrophoresis-mass spectrometry as a useful tool for highly efficient and high-capacity separation of proteoforms and protein complexes as well as the combination of electron capture dissociation and collision-induced dissociation as a powerful technique for nearly 100% backbone cleavage of proteins smaller than 30 kDa. We employed an Agilent G1700 Capillary Electrophoresis with CMP EMass II interface, 6545XT Q-TOF mass spectrometer and eMSion ECD cell in these studies.

Sun, Liangliang PIC

Dr. Liangliang Sun is an Assistant Professor in the Department of Chemistry at Michigan State University. He joined MSU in August 2016. Before that, he worked with Prof. Norman Dovichi at University of Notre Dame as a postdoctoral fellow (2011-2012) and a Research Assistant Professor (2013-2016). He received his Ph.D. degree in Analytical Chemistry in 2011 from Dalian Institute of Chemical Physics, Chinese Academy of Sciences, advised by Profs. Yukui Zhang and Lihua Zhang. He got his bachelor degree in Bioengineering in 2005 from Dalian University of Technology, China.

Pictures for bottom of emails - 5 pics going across