Development of an ECD cell for biomolecular characterization
EMSL Project ID
50157
Abstract
The ultimate goal of this proposal is to integrate an electron capture dissociation (ECD) cell into ThermoFisher Scientific’s Exactive series mass spectrometers. The proposed work in collaboration with eMSion will enable design and development of the ECD cell and subsequent evaluation of ECD performance for standard bottom-up and top-down proteomics experiments. EMSL is uniquely capable of performing this work due a close collaboration with ThermoFisher Scientific that allows access to the firmware controlling the Exactive mass spectrometers. This capability will enable evaluation and testing of the ECD cell that is not possible anywhere else outside of the manufacturer. The capability to perform ECD in addition to other existing tandem mass spectrometry methods will enhance polypeptide de novo sequencing and native mass spectrometry capabilities at the EMSL. Methodologies developed in the work will be directly applicable to ongoing DOE funded work to characterized microbial secreted enzymes responsible for organic matter degradation.
Project Details
Start Date
2018-02-08
End Date
2020-04-29
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Shaw J.B., N. Malhan, Y.V. Vasilev, N.I. Lopez, A. Makarov, J.S. Beckman, and V.G. Voinov. 2018. "Sequencing Grade Tandem Mass Spectrometry for Top-Down Proteomics Using Hybrid Electron Capture Dissociation Methods in a Benchtop Orbitrap Mass Spectrometer." Analytical Chemistry 90, no. 18:10819-10827. PNNL-SA-133760. doi:10.1021/acs.analchem.8b01901
Shaw J.B., W. Liu, Y.V. Vasil'Ev, C.C. Bracken, N. Malhan, A.L. Guthals, and J.S. Beckman, et al. 2020. "Direct Determination of Antibody Chain Pairing by Top-down and Middle-down Mass Spectrometry using Electron Capture Dissociation and Ultraviolet Photodissociation." Analytical Chemistry 92, no. 1:766-773. PNNL-SA-145157. doi:10.1021/acs.analchem.9b03129
Zhou M., W. Liu, and J.B. Shaw. 2020. "Charge Movement and Structural Changes in the Gas-Phase Unfolding of Multimeric Protein Complexes Captured by Native Top-Down Mass Spectrometry." Analytical Chemistry 92, no. 2:1788-1795. PNNL-SA-145158. doi:10.1021/acs.analchem.9b03469