Title
Sizing up large protein complexes by electrospray ionisation-based electrophoretic mobility and native mass spectrometry: Morphology selective binding of Fabs to hepatitis B virus capsids
Author
Bereszczak, J.Z.
Havlik, M.
Weiss, V.U.
Marchetti-Deschmann, M.
van Duijn, E.
Watts, N.R.
Wingfield, P.T.
Allmaier, G.
Steven, A.C.
Heck, A.J.R.
Publication year
2014
Abstract
The capsid of hepatitis B virus (HBV) is a major viral antigen and important diagnostic indicator. HBV capsids have prominent protrusions ('spikes') on their surface and are unique in having either T = 3 or T = 4 icosahedral symmetry. Mouse monoclonal and also human polyclonal antibodies bind either near the spike apices (historically the 'α-determinant') or in the 'floor' regions between them (the 'β-determinant'). Native mass spectrometry (MS) and gas-phase electrophoretic mobility molecular analysis (GEMMA) were used to monitor the titration of HBV capsids with the antigen-binding domain (Fab) of mAb 3120, which has long defined the β-determinant. Both methods readily distinguished Fab binding to the two capsid morphologies and could provide accurate masses and dimensions for these large immune complexes, which range up to ~8 MDa. As such, native MS and GEMMA provide valuable alternatives to a more time-consuming cryo-electron microscopy analysis for preliminary characterisation of virus-antibody complexes. [Figure not available: see fulltext.] © 2013 Springer-Verlag Berlin Heidelberg.
Subject
Life
MSB - Microbiology and Systems Biology
ELSS - Earth, Life and Social Sciences
Biomedical Innovation
Biology
Healthy Living
GEMMA
Hepatitis B
Immune complex
Native MS
Quasi-equivalence
Virus-antibody complexes
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http://resolver.tudelft.nl/uuid:394a863a-c9df-4d33-8ca2-141bb7c84ca8
DOI
https://doi.org/10.1007/s00216-013-7548-z
TNO identifier
492977
ISSN
1618-2642
Source
Analytical and Bioanalytical Chemistry, 406 (5), 1437-1446
Document type
article