DOI: 10.1101/501908Dec 19, 2018Paper

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

BioRxiv : the Preprint Server for Biology
Christopher S HughesGregg B Morin

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 nano-chromatography columns with larger IDs, substantially improved throughput with no reduction in detection sensitivity is achieved in single-injection proteome analyses. 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...Continue Reading

Related Concepts

Chromatography
Liquid Chromatography
Spectrometry, X-Ray Emission
LC 6
Cohort
Proteome
Proteomics
Analysis
Proteomic Profiling
Detection

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