期刊
ANALYTICAL CHEMISTRY
卷 91, 期 12, 页码 7510-7515出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.9b00118
关键词
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资金
- Deutsche Forschungsgemeinschaft [INST 371/23-1 FUGG, SFB1066 B11, TE599/3-1, SFB1292 Z1, DI 2471/1-1]
- Forschungszentrum Immunologie (FZI) of the Johannes Gutenberg University Mainz
- Forschungszentrum Translationale Neurowissenschaften (FTN) of the Johannes Gutenberg University Mainz
The introduction of more sensitive mass spectrometers allows researchers to adapt front-end liquid chromatography (LC) to individual needs for the analysis of complex proteomes. Where absolute sensitivity is not paramount, it is advantageous to switch from a highly sensitive nanoflow-LC setup, the de facto standard platform in mass spectrometry (MS)-based discovery proteomics, to a more robust, high-throughput-compatible microflow or conventional-flow setup. To enhance the microflow-LC-MS electrospray process of complex proteomic samples, we tested the effects of different solvents, including 2-propanol, methanol, and acetonitrile, pure or as mixture with dimethyl sulfoxide, which were added postcolumn to the eluting sample. Postcolumn addition of organic solvents strongly enhanced the electrospray efficiency in microflow-LC-MS experiments and improved the sensitivity across the entire gradient and for early eluting peptides by up to 10-fold. Postcolumn solvent addition did not negatively affect chromatographic performance and resulted in an overall 28-36% increase in identifications at both the protein and peptide levels. The presented microflow-LC-MS workflow, including postcolumn solvent addition, can be easily adopted on any LC-MS/MS platform.
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