April 30-May 2, 2018 : Targeted proteomics with Skyline
Description
This 2.5 day short course will walk through the process of designing, developing, and statistically evaluating LC-MS methods for the targeted quantification of peptides with high confidence. Topics covered will include peptide selection from target proteins, optimization of peptide transitions, LC gradient, collision energy, and discussion on internal standards. Attendees will learn rigorous method evaluation and performance characterization, limits of detection and quantification calculation, and use of routine System Suitability QC. Case studies and tutorials will be provided for hands-on participation with open-source software Skyline (https://skyline.ms/project/home/software/Skyline/begin.view).
Target audience
- Target audience are experimental scientists with experience in proteomic research. We also welcome computational scientists interested in working with raw mass spectrometric data.
Speakers
- Sue Abbatiello, Andy Hoofnagle, Brendan MacLean, Lindsay Pino
Tentative schedule
Monday, April 30, 2018
- 8:00 a.m. Registration
- 9:00 a.m. Keynote : General peptide Quan/CPTAC, Susan Abbatiello
- 10:00 a.m. Introduction to Skyline, Brendan MacLean
- 10:30 a.m. Refreshments
- 11:00 a.m. Hands-on : Developing a targeted list, refinding transitions, optimizing collision energy, Susan Abbatiello
- 11:45 a.m. iRT, retention time, standards, Lindsay Pino
- 12:30 p.m. Lunch
- 1:30 p.m. Hands-on : Plasma dataset, Lindsay Pino
- 2:00 p.m. LC optimization, Lindsay Pino
- 3:00 p.m. Refreshments
- 3:30 p.m. Hands-on: iRT calculator, Brendan MacLean
- 5:00 p.m. Q&A and adjourn
Tuesday, May 1, 2018
- 8:00 a.m. Skyjam with Brendan
- 9:00 a.m. Keynote : HDL Proteomics in Chronic Kidney Disease, Andy Hoofnagle
- 10:30 a.m. Refreshments
- 11:00 a.m. Standards, Peptide stability, reproducibility, repeatability, Lindsay Pino
- 12:30 p.m. Lunch
- 1:30 p.m. Absolute quantification and method validation: Calibration curves, Susan Abbatiello
- 3:00 p.m. Refreshments
- 3:30 p.m. Experimental design for a curve, Susan Abbatiello
- 4:00 p.m. LOD/LOQ methods comparison, Lindsay Pino
- 4:30 p.m. Review of Skyline Ecosystem, Lindsay Pino
- 5:00 p.m. Q&A and adjourn
Wednesday, May 2, 2018
- 8:00 a.m. Skyjam with Brendan
- 9:00 a.m. System suitability, QC design, Susan Abbatiello
- 10:30 a.m. Refreshments
- 11:00 a.m. System suitability evaluation, Lindsay Pino
- 12:30 p.m. Wrap-up and Kahoot!