Speaker: Kyle Anderson, National Institute of Standards and Technology

Topic: Advancements in methods and instrumentation for HDX-MS and LC-MS

Date: January 13, 2025

Time: 6:15 pm Dinner, 7:15 pm Presentation

Location: Shimadzu Scientific Instrument, Inc. Training Center 7100 Riverwood Drive, Columbia, MD 21046 (Directions)

Dinner: Please RSVP to Dingyin Tao (owendtao@gmail.com) by Friday, January 10 if you will be attending the dinner.

Abstract: Subzero temperature chromatography minimizes loss of deuterium during HDX-MS analysis. We developed an HDX-MS chromatography apparatus capable of performing long analytical separations at -30 °C. The apparatus has precise temperature control with two enzyme column compartments held at independent temperatures and allows for rigorous cleaning in parallel to data acquisition to reduce instrument downtime and sample carryover. Methods for both reversed phase and HILIC analytical separations at subzero temperature were developed. HILIC methods developed greatly reduced operational backpressures for easier adoption using any HPLC pumps and enabled virtually water-free separations at subzero temperature. An HPLC pump delivering custom wash solutions to immobilized protease columns was added to replace conventional injections of common wash solutions, which greatly reduce sample carryover and cut necessary blank runs from 5 to 1 for an IgG1 digest. Additionally, methods for automated online removal of phospholipids from membrane proteins will be presented.

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Lightning Talk
Jacob Epstein (NIH/NIA/IRP)