Top-down mass spectrometry of native proteoforms and their complexes: A community study

Area of Science:

• Proteomics
• Mass Spectrometry
• Biochemistry

Background:

• Native electrospray ionization coupled with top-down fragmentation enables simultaneous analysis of noncovalent complex stoichiometry, proteoforms, and co-factors.
• The adoption of native and top-down mass spectrometry is limited due to a lack of standardized protocols.

Approach:

• The Consortium for Top-Down Proteomics (CTDP) conducted a multi-laboratory study involving eleven instruments across nine institutions.
• Protocols for native mass spectrometry combined with top-down fragmentation of proteins and protein complexes were developed and rigorously tested.

Key Points:

• The study aimed to establish standardized methodologies for native and top-down mass spectrometry.
• Eleven instruments across nine laboratories participated in the protocol testing.
• The research focused on analyzing protein complexes and their components.

Conclusions:

• This report summarizes the outcomes of the protocol development and testing phase.
• The findings provide robust benchmarks for the scientific community.
• The study serves as a valuable entry point for researchers new to these advanced mass spectrometry techniques.