High-flow Nano-chromatography Columns Facilitate Rapid and High-quality Proteome Analysis on Standard Nano-LC Hardware

Abstract

Optimizing the acquisition of proteomics data collected from a mass spectrometer (MS) requires careful selection of processed material quantities, liquid-chromatography (LC) setup, and data acquisition parameters. The small internal diameter (ID) columns standardly used in nano-chromatography coupled MS result in long per injection overhead times that require sacrifices in design of offline-fractionation and data acquisition schemes. As cohort sizes and the numbers of samples to be analyzed continue to increase, there is a need to investigate methods for improving the efficiency and time of an acquisition (LC + MS). In this work, the ability to improve throughput in single runs or as part of an in-depth proteome analysis of a fractionated sample using standard LC hardware is investigated. Capitalizing on the increased loading capacity of nanochromatography columns with larger IDs, substantially improved throughput with no reduction in detection sensitivity is achieved in single-injection proteomeanalyses. An optimized 150 μm ID column setup is paired with an offline fractionation-concatenation scheme to demonstrate the ability to perform in-depth proteome analysis on-par with current state-of-the-art studies, while minimizing sample loading overhead. Together, these data demonstrate an easy and effective means to improve sample analysis throughput with no reduction in data quality using an approach that is applicable to any standard nano-LC hardware.3

Electroactive Polymer-Based Spray Ionization for Direct Mass Spectrometric Analysis

“…All MS experiments were carried out using a Thermo Scientific Velos Pro dual-cell linear ion trap mass spectrometer fitted with an entirely custom ionization source based on the University of Washington’s design . A mass range of m / z 150–1000 was used to analyze CBD and CBN, while other analyses used a mass range of m / z 150–2000.…”

“…All MS experiments were carried out using a Thermo Scientific Velos Pro dual-cell linear ion trap mass spectrometer fitted with an entirely custom ionization source based on the University of Washington's design. 51 A mass range of m/z 150−1000 was used to analyze CBD and CBN, while other analyses used a mass range of m/z 150−2000. In the EAPSI ionization source (Figure 1b,c), a positioner was set up in front of the sample cone, which holds the substrate surface, i.e., the PEDOT-C 12 coated ITO glass slide or a piece of paper.…”