Poster Presentations
Day 3, June 12（Fri.） Room P (5F 501+502)
- 3P-30
Increasing Throughput in Collision-Induced Unfolding Studies: Enhancing Sensitivity and Streamlining Set-up
(1Nihon Waters, 2Waters)
oMaki Terasaki1, Etsuko Yada1, Cooper-Shepherd Dale2, Marsden-Edwards Emma2, Kenji Hirose1
Collision-induced unfolding (CIU) is a valuable technique for the study of the structure and stability of biomolecules and their complexes. Tracking discrete conformational states on the unfolding pathway enables the evaluation of domain stability, ligand binding and proteoform- or mutation-dependent effects. In this work we describe two innovations implemented on a Cyclic IMS; firstly, we have introduced a CIU acquisition mode in the method editor, making set-up rapid and simple and secondly, we enable an enhanced duty cycle mode of acquisition to improve sensitivity and reduce run times. As a proof-of-principle, we selected Streptavidin as a model system. All data were acquired after direct infusion. Data were processed using CIUSuite2. We created a CIU method for the 16+ charge state with 15steps from 5 V to 75 V. The resulting CIU unfolding profile shows four major conformational families from folded to unfolded. Next, we investigated the impact of TOF duty cycle increases on the intensity of the signals. In this case, we observed a signal increase of three times versus the unenhanced condition. Further investigations into signal gains will be presented as well as data from a monoclonal antibody.