Signature of Antibody Domain Exchange by Native Mass Spectrometry and Collision-Induced Unfolding.

TitleSignature of Antibody Domain Exchange by Native Mass Spectrometry and Collision-Induced Unfolding.
Publication TypeJournal Article
Year of Publication2018
AuthorsWatanabe Y, Vasiljevic S, Allen JD, Seabright GE, Duyvesteyn HME, Doores KJ, Crispin M, Struwe WB
JournalAnal Chem
Volume90
Issue12
Pagination7325-7331
Date Published06/19/2018
ISSN1520-6882
Abstract

The development of domain-exchanged antibodies offers a route to high-affinity targeting to clustered multivalent epitopes, such as those associated with viral infections and many cancers. One strategy to generate these antibodies is to introduce mutations into target antibodies to drive domain exchange using the only known naturally occurring domain-exchanged anti-HIV (anti-human immunodeficiency virus) IgG1 antibody, 2G12 , as a template. Here, we show that domain exchange can be sensitively monitored by ion-mobility mass spectrometry and gas-phase collision-induced unfolding. Using native 2G12 and a mutated form that disrupts domain exchange such that it has a canonical IgG1 architecture ( 2G12 I19R ), we show that the two forms can be readily distinguished by their unfolding profiles. Importantly, the same signature of domain exchange is observed for both intact antibody and isolated Fab fragments. The development of a mass spectrometric method to detect antibody domain exchange will enable rapid screening and selection of candidate antibodies engineered to exhibit this and other unusual quaternary antibody architectures.

DOI10.1021/acs.analchem.8b00573
Alternate JournalAnal. Chem.
PubMed ID29757629
PubMed Central IDPMC6008249
Grant ListUM1 AI100663 / AI / NIAID NIH HHS / United States
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