In addition to mass spectrometry techniques, the choice of an appropriate sample preparation method is critical for top-down proteomics. But there are a variety of sample preparation methods for intact proteoforms. Which one should you use? Dr. Kaulich gave a very clear answer to this difficult question at this year’s ASMS conference in Anaheim just a few weeks ago. Join us for this webinar to learn about the best sample preparation methods available today!

References for this lecture

Influence of Different Sample Preparation Approaches on Proteoform Identification by Top-Down Proteomics.

Kaulich PT, Jeong K, Kohlbacher O, Tholey A.

PREPRINT (Version 1) available at Research Square. 2024 Mar 20. https://doi.org/10.21203/rs.3.rs-3990966/v1.

Improved Identification of Proteoforms in Top-Down Proteomics Using FAIMS with Internal CV Stepping.

Kaulich PT, Cassidy L, Winkels K, Tholey A.

Anal Chem. 2022 Mar 1;94(8):3600-3607. doi: 10.1021/acs.analchem.1c05123.

"I'm interested in top-down proteomics, but I’ll need special mass spectrometry equipment, right?" This is a question we often get from JPrOS members. Dr. Fornelli is a true expert who has successfully analyzed many intact proteoforms using bench-top Orbitrap mass spectrometers which many of you already have access to. In his talk, you will get insights into new proteoform analysis techniques using state-of-the-art mass spectrometry, as well as practical advice on how to start top-down proteomics studies in your own laboratory.

References for this lecture

Targeted Quantification of Proteoforms in Complex Samples by Proteoform Reaction Monitoring.

Huang CF, Kline JT, Negrão F, Robey MT, Toby TK, Durbin KR, Fellers RT, Friedewald JJ, Levitsky J, Abecassis MMI, Melani RD, Kelleher NL, Fornelli L.

Anal Chem. 2024 Feb 27;96(8):3578-3586. doi: 10.1021/acs.analchem.3c05578.

Orbitrap Mass Spectrometry and High-Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS) Enable the in-Depth Analysis of Human Serum Proteoforms.

Kline JT, Belford MW, Boeser CL, Huguet R, Fellers RT, Greer JB, Greer SM, Horn DM, Durbin KR, Dunyach JJ, Ahsan N, Fornelli L.

J Proteome Res. 2023 Nov 3;22(11):3418-3426. doi: 10.1021/acs.jproteome.3c00488.

To accurately identify intact proteoforms by mass spectrometry, it is important not only to obtain high-resolution MS information from large protein molecules, but also to dissociate their ions into fragments for determining their post-translational modifications and sequences. Dr. Baba will give a highly informative lecture covering everything from the principles of proteoform identification by mass spectrometry to the latest dissociation methods for top-down proteomics.

References for this lecture

Analysis of Sialic Acid Linkage in N-Linked Glycopeptides Using Liquid Chromatography-Electron-Activated Dissociation Time-of-Flight Mass Spectrometry.

Liu S, Ryumin P, Albanese J, Zhang Z, Baba T.

Anal Chem. 2023 May 16;95(19):7458-7467. doi: 10.1021/acs.analchem.2c04581.

Localization of Multiple O-Linked Glycans Exhibited in Isomeric Glycopeptides by Hot Electron Capture Dissociation.

Baba T, Zhang Z, Liu S, Burton L, Ryumin P, Le Blanc JCY.

J Proteome Res. 2022 Oct 7;21(10):2462-2471. doi: 10.1021/acs.jproteome.2c00378.