Program

Poster Presentations

Day 2, June 11（Thu.）　　Room P (5F 501+502)

2P-46

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Development of a General Method Using Multi‑turn TOF‑MS for High‑Resolution Mass Analysis on Intact Proteins and Noncovalent Complexes

(¹Shimadzu, ²UOsaka)
^oQiuyi Wang¹, Yusuke Tateishi¹, Hiroyuki Miura¹, Hiroko Morinaga¹, Yoshinori Arita¹, Masaru Nishiguchi¹, Zi Wang², Toshifumi Takao², Daisuke Okumura¹

Large proteins (e.g., enzymes, antibodies) show high heterogeneity from multi domain architecture, complex glycosylation, diverse PTMs and noncovalent interactions with metals or small ligands. This affects function, drug stability and immunogenicity; therefore high resolution mass analysis of intact proteins is needed to distinguish proteoforms, detect small mass differences, and assess coordination states. Conventional MS often fails for high mass species: ionization/transmission efficiencies fall with mass, reducing sensitivity, while peak broadening and limited resolving power hinder separation of closely spaced proteoforms, especially when preserving noncovalent interactions.
We employed a multi turn time of flight (multi-turn TOF) method and developed a generalized analysis workflow. The instrument confines ions on extended 3D orbits for multiple revolutions, markedly enhancing mass resolution in a compact footprint; prior reports show resolving power > 200,000 for high mass analytes. We implemented gentle volatile buffer exchange to ammonium acetate (SEC or MWCO dialysis), nano electrospray ionization, and optimization of key TOF parameters to form a broadly applicable acquisition protocol. Representative enzymes, antibodies and transferrin validated generality. The method enables high precision mass measurement and separation of closely spaced proteoforms while largely preserving noncovalent interactions and intact conformations.

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