학술논문

Standard Proteoforms and Their Complexes for Native Mass Spectrometry.
Document Type
Article
Source
Journal of the American Society for Mass Spectrometry. Jul2019, Vol. 30 Issue 7, p1190-1198. 9p.
Subject
*MASS spectrometry
*PROTEOMICS
*PROTEINS
*ANALYTICAL chemistry
*CHEMICAL sample preparation
Language
ISSN
1044-0305
Abstract
Native mass spectrometry (nMS) is a technique growing at the interface of analytical chemistry, structural biology, and proteomics that enables the detection and partial characterization of non-covalent protein assemblies. Currently, the standardization and dissemination of nMS is hampered by technical challenges associated with instrument operation, benchmarking, and optimization over time. Here, we provide a standard operating procedure for acquiring high-quality native mass spectra of 30–300 kDa proteins using an Orbitrap mass spectrometer. By describing reproducible sample preparation, loading, ionization, and nMS analysis, we forward two proteoforms and three complexes as possible standards to advance training and longitudinal assessment of instrument performance. Spectral data for five standards can guide assessment of instrument parameters, data production, and data analysis. By introducing this set of standards and protocols, we aim to help normalize native mass spectrometry practices across labs and provide benchmarks for reproducibility and high-quality data production in the years ahead. [ABSTRACT FROM AUTHOR]