A Simple and Flexible Infusion Platform for Automated Native Mass Spectrometry Analysis

Abstract

High throughput native mass spectrometry analysis of proteins and protein complexes has been enabled by recent development of infusion and liquid chromatography (LC) systems, which often include complete LC pumps without fully utilizing their gradient flows. We demonstrated a lower-cost infusion cart for native mass spectrometry applications using a single isocratic solvent pump that can operate at both nano-and high-flow configurations (0.05−150 μL/min) for both infusion and online buffer exchange experiments. The platform is controlled via opensource software and can potentially be expanded for customized experimental designs, offering a lower cost alternative to laboratories with limited budgets and/or needs in student training.

Laser capture microdissection and native mass spectrometry for spatially-resolved analysis of intact protein assemblies in tissue

“…Our findings suggest that LCMD provides a complementary approach to nano-DESI and LESA (both with integrated and decoupled ionisation) for identification of proteins detected in native ambient mass spectrometry imaging. Future work could focus on improving throughput by transferring the dissected regions into a well plate and use of a native MS infusion cart 36 to automate the MS analysis.…”

Expanding Native Mass Spectrometry to the Masses

“…More recently a simple infusion platform using a single isocratic pump has been presented for use with online buffer exchange and offers a lower cost alternative to an LC for such studies. 6 Online desalting approaches remain the highest throughput and would have the greatest appeal to those wishing to create a walk-up or open access facility.…”

“…A hurdle to online approaches, particularly within a core facility or to those just beginning in the field, can be the requirement for an LC, which can sometimes be cost prohibitive. More recently a simple infusion platform using a single isocratic pump has been presented for use with online buffer exchange and offers a lower cost alternative to an LC for such studies . Online desalting approaches remain the highest throughput and would have the greatest appeal to those wishing to create a walk-up or open access facility.…”

Native Mass Spectrometry Dissects the Structural Dynamics of an Allosteric Heterodimer of SARS-CoV-2 Nonstructural Proteins

“…Native mass spectrometry (native MS) characterizes noncovalent complexes under nondenaturing conditions and can provide fast assessment (typically in minutes) of protein–protein and protein–ligand interactions using small quantities (∼μg) of often precious samples. − Although native MS requires specialized methods distinct from common MS applications, recent advances in commercial instrumentation and automated systems have significantly increased the accessibility of the technology. − The methodologies for studying protein–ligand interactions have been reviewed in detail recently . However, most applications focus on the intact mass measurement at the MS1 level.…”