Speaker: Andy Y. Qi, National Institutes of Health
Topic: A fully automated FAIMS-DIA proteomic pipeline for high-throughput characterization of iPSC-derived neurons
Date: Monday, December 13, 2021
Time: 6:45 Virtual Social, 7:15 pm Presentation
Location: See Zoom invite in email on December 2 and 10.
Abstract: Fully automated proteomic pipelines have the potential to achieve deep coverage of cellular proteomes with high throughput and scalability. However, it is important to evaluate performance, including both reproducibility and ability to provide meaningful levels of biological insight. Here, we present an approach combining high field asymmetric waveform ion mobility spectrometer (FAIMS) interface and data independent acquisition (DIA) proteomics approach developed as part of the induced pluripotent stem cell (iPSC) Neurodegenerative Disease Initiative (iNDI), a large-scale effort to understand how inherited diseases may manifest in neuronal cells. Our FAIMS-DIA approach identified more than 8000 proteins per mass spectrometry (MS) acquisition as well as superior total identification, reproducibility, and accuracy compared to other existing DIA methods. Next, we applied this approach to perform a longitudinal proteomic profiling of the differentiation of iPSC-derived neurons from the KOLF2.1J parental line used in iNDI. This analysis demonstrated a steady increase in expression of mature cortical neuron markers over the course of neuron differentiation. We validated the performance of our proteomics pipeline by comparing it to single cell RNA-Seq datasets obtained in parallel, confirming expression of key markers and cell type annotations. An interactive webapp of this temporal data is available for aligned-UMAP visualization and data browsing (https://share.streamlit.io/anant-droid/singlecellumap). In summary, we report an extensively optimized and validated proteomic pipeline that will be suitable for large-scale studies such as iNDI.
Topic: Post-ASMS Poster Night and ASMS Travel Award Presentations
Date: Monday, November 15, 2021
Time: 6:15 pm Dinner (outdoors) and Vendor Night, 7:30 pm Presentations
Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)
This will be an in-person meeting. Attendees are required to show a vaccine card (either at the door or in advance using the web form) and to wear a mask.
Dinner: Please RSVP to Dapeng Chen (firstname.lastname@example.org) by Friday, November 12th if you will be attending the dinner and/or if you will participate in the poster session.
ASMS Travel Award Recipients:
- Anh Tran, University of Maryland School of Pharmacy
: “High Resolution Ion Mobility of Sphingolipids: a Multi-Adduct Perspective”
- Aparna Baxi, University of Maryland
: “In “Proteo-Metabolomics of Spemann’s Organizer in the Vertebrate (Frog) Embryo”
- Jie Li, University of Maryland
: “In Vivo Subcellular Mass Spectrometry Enables Systems Biology in Single Embryonic Cells”
- Amanda Belunis, University of Maryland Baltimore County
: “EPA 537.1 method validation for the detection of per- and polyfluoroalkyl substances (PFAS) in drinking water sources”
Congratulations to the following recipients of the WBMSDG Young Investigator Travel Awards:
Anh Tran, University of Maryland School of Pharmacy (Advisor: Jace Jones), 1st place: “High Resolution Ion Mobility of Sphingolipids: a Multi-Adduct Perspective”
Aparna Baxi & Jie Li, University of Maryland (Advisor: Peter Nemes), 2nd place: “In “Proteo-Metabolomics of Spemann’s Organizer in the Vertebrate (Frog) Embryo” and “In Vivo Subcellular Mass Spectrometry Enables Systems Biology in Single Embryonic Cells”
Amanda Belunis, University of Maryland Baltimore County (Advisor: William R. LaCourse), 3rd place: “EPA 537.1 method validation for the detection of per- and polyfluoroalkyl substances (PFAS) in drinking water sources”
Speaker: Peter Nemes, University of Maryland
Topic: Mass Spectrometry for Discovering the Cell
Date: Monday, October 18, 2021
Time: 6:45 Virtual Social, 7:15 pm Presentation
Location: MOVED ONLINE. See Zoom invite in email on October 14.
Abstract: Knowledge of all the types of molecules that are produced in cells as they establish different tissues and organs is key to understanding normal development and design efficient therapeutics. Even today, after the sequencing of entire genomes, there is limited information on how molecules downstream, such as proteins and metabolites, contribute to cell processes. The limitation has been a lack of sufficiently sensitive mass spectrometry technologies that can measure these biomolecules with scalability in space and time and compatibility for live development, a prerequisite for functional biology. In this presentation, we will discuss technological developments from our laboratory to to transform mass spectrometry proteomics and metabolomics to single cells. We used these technologies to determine the proteomic and metabolomic profile of identified cells in live Xenopus laevis frog embryos and neurons in mouse brain tissues. Molecular measurements with separation using capillary electrophoresis and detection by (trapped ion mobility) time-of-flight or orbitrap mass spectrometry revealed quantitative molecular differences between cells of different phenotypes. Through follow-up functional experiments, we discovered molecules capable of altering normal cell fate decisions in the chordate embryo. The technology was scalable to smaller cells, including electrophysiologically identified neurons in the mouse brain. Quantification of ~300–800 different proteins revealed reproducible proteomic differences between dopaminergic, serotonergic, and parvalbumin interneurons. Microprobe capillary electrophoresis mass spectrometry expands the molecular toolbox of cell biology and neuroscience.