期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 61, 期 25, 页码 -出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202115047
关键词
Cold Denaturation; Monoclonal Antibodies; Protein Folding; Structure-Activity Relationships; Variable-Temperature Ion Mobility
资金
- Research Impact Scholarship - alumni of the University of Manchester
- Bristol-Myers Squibb
- BBSRC Industrial Case Studentship
- Leverhulme Trust [RPG-2019-178]
- EPSRC [EP/T019328/1, GR/S77639/01]
- BBSRC [BB/L01646X/1 BB/L015048/1 BB/L002655/1]
- Covance Laboratories
This study investigates the conformational changes of antibodies at different temperatures using variable-temperature ion mobility mass spectrometry. It is found that at lower temperatures, the structure of the antibodies does not rearrange, while within a certain temperature range, the changes in the strength of intermolecular interactions lead to restructuring.
The effect of temperature on the stability of proteins is well explored above 298 K, but harder to track experimentally below 273 K. Variable-temperature ion mobility mass spectrometry (VT IM-MS) allows us to measure the structure of molecules at sub-ambient temperatures. Here we monitor conformational changes that occur to two isotypes of monoclonal antibodies (mAbs) on cooling by measuring their collision cross sections (CCS) at discrete drift gas temperatures from 295 to 160 K. The CCS at 250 K is larger than predicted from collisional theory and experimental data at 295 K. This restructure is attributed to change in the strength of stabilizing intermolecular interactions. Below 250 K the CCS of the mAbs increases in line with prediction implying no rearrangement. Comparing data from isotypes suggest disulfide bridging influences thermal structural rearrangement. These findings indicate that in vacuo deep-freezing minimizes denaturation and maintains the native fold and VT IM-MS measurements at sub ambient temperatures provide new insights to the phenomenon of cold denaturation.
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