A word or two about our speakers and their lectures
Dr. Liangliang Sun is an Assistant Professor in the Department of Chemistry at Michigan State University (East Lansing, Michigan, USA). He received his Ph.D. in Analytical Chemistry from Dalian Institute of Chemical Physics, Chinese Academy of Sciences (Dalian, China) in 2011. Then he worked with Prof. Norman Dovichi as a postdoctoral fellow and later as a research assistant professor from 2011 to 2016 at the University of Notre Dame (Notre Dame, Indiana, USA). He joined Michigan State University as an Assistant Professor in 2016. He has 12-years’ experiences on proteomics and has published 84 peer-reviewed papers with over 1700 citations.
Dr. Sun’s research group at MSU aims to develop novel analytical methodologies based on capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) for comprehensive and highly sensitive bottom-up and top-down proteomics. His research group applies those methodologies for quantitative proteomics of zebrafish embryos and blastomeres during early embryogenesis to achieve the dynamics of gene expression and protein post-translational modifications (PTMs) during early embryogenesis at high time resolution and single-cell resolution
Top-down Proteomics Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry
Genome-level and transcriptome-level information cannot accurately reflect proteome-level information because post-transcriptional regulations modulate gene expression, because protein post-translational modifications (PTMs) influence protein function, and because most proteins in cells function as complexes with proteins, RNAs, metals or other small molecules. Characterization of the proteome is imperative to understand the roles played by proteins, protein PTMs and even protein complexes in development and diseases.
Top-down proteomics is a well-known strategy for large-scale characterization of proteome at the intact protein level and is very useful for high-resolution characterization of proteoforms that represent all kinds of protein molecules derived from the same gene due to gene-level variations, RNA-level alternative splicing, and protein-level PTMs. The number of proteoforms in the human proteome has been estimated to be over 1 million. High-capacity separation of proteoforms before electrospray ionization-mass spectrometry and tandem mass spectrometry (ESI-MS and MS/MS) is essential for deep and high-resolution top-down proteomics.
The top-down proteomics community has made tremendous efforts in improving liquid chromatography (LC)-MS and MS/MS for top-down proteomics. We argue that capillary zone electrophoresis (CZE)-MS and MS/MS is another powerful tool for top-down proteomics because CZE can approach high-capacity separation of intact proteins and because CZE-MS has shown obviously higher sensitivity than LC-MS for protein detection. In my talk, I will talk about our recent work on boosting CZE-MS and MS/MS for large-scale and highly sensitive top-down proteomics.[4,5] I will also introduce our work on developing CZE-MS and MS/MS methodologies for native top-down proteomics that aims to characterize the endogenous protein complexes in cells in discovery mode.
 Smith et al. Nat Methods 2013, 10, 186-187.
 Aebersold et al. Nat Chem Biol. 2018, 14, 206-214.
 Han et al. J Proteome Res. 2014, 13, 6078-6086.
 Lubeckyj et al. Anal Chem. 2017, 89, 12059-12067.
 McCool et al. Anal Chem. 2018, 90, 5529-5533.
 Shen et al. Anal Chem. 2018, 90, 10095-10099.
William Farrell is Associate Research Fellow at Pfizer Global R&D - La Jolla. Mr. Farrell received his BS degree from SUNY Stony Brook and has over 25 years chromatographic experience, specializing in supercritical fluid (SFC) and high pressure liquid (HPLC) chromatographic techniques. He has spent the last 20 years working on the implementation of SFC and HPLC for the analysis & purification of medicinal and combinatorial derived compounds. He has been a major driving force behind the development of automated Preparative SFC systems and has pioneered the use of SFC for high throughput applications for which he received the first annual Averica Award for his contributions to Preparative SFC and the ACS Technical Achievement in Organic Chemistry (TAOC) award. Current research areas include expanding SFC applications into biological systems and the development of novel, SFC specific stationary phases.
Supercritical Fluid Chromatography Mass Spectrometry Impact on Medicinal Chemistry Applications
Faced with a separation challenge, most chemists would reach for a reversed phase HPLC column and apply a generic gradient from 100% aqueous to full organic. The reason is likely because the majority of these chromatographers have built upon their own personal experience with RP-HPLC, either in academia or industry, using this approach as a starting point. The simplicity of the setup, broad applicability and the relative ease to learn has made RP-HPLC a popular, near universal technique. With recent advanced in smaller particles and more sophisticated instrumentation, the technique has only gained in popularity. However, RP-HPLC does have some limitations, especially when retaining polar compounds or samples that contain a mix of compounds with a large span of Log D's, which make method development challenging. These types of samples have led to the resurgence in Hydrophilic Interaction Liquid Chromatography (HILIC), an aqueous based, normal phase technique, which uses polar stationary phases with highly organic mobile phases.
This presentation will highlight developments in Supercritical Fluid technology that have opened new opportunities to impact Medicinal Chemistry projects. One example will describe how high throughput synthetic screening efficiency and data analysis quality can be exponentially improved through the use of customized software solutions. In addition, highlights of the utilization of SFC/MS technology combined with unique stationary phases to fully resolve, identify and quantitate reaction mixtures which were otherwise intractable using RP-HPLC/ESI-MS (the “gold standard”) will be presented.