Binding symmetry and surface flexibility mediate antibody self-association

From National Research Council Canada

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DOI	Resolve DOI: https://doi.org/10.1080/19420862.2019.1632114
Author	Search for: Schrag, Joseph D.¹; Search for: Picard, Marie-Ève ORCID identifier: https://orcid.org/0000-0002-4316-2830; Search for: Gaudreault, Francis¹ORCID identifier: https://orcid.org/0000-0001-5656-3118; Search for: Gagnon, Louis-Patrick¹ORCID identifier: https://orcid.org/0000-0001-7417-866X; Search for: Baardsnes, Jason¹ORCID identifier: https://orcid.org/0000-0001-5587-3034; Search for: Manenda, Mahder S.ORCID identifier: https://orcid.org/0000-0002-8640-5556; Search for: Sheff, Joey¹ORCID identifier: https://orcid.org/0000-0002-9269-6863; Search for: Deprez, Christophe¹ORCID identifier: https://orcid.org/0000-0003-2687-3144; Search for: Baptista, Cassio¹; Search for: Hogues, Hervé¹; Search for: Kelly, John F.¹; Search for: Purisima, Enrico O.¹; Search for: Shi, Rong ORCID identifier: https://orcid.org/0000-0001-8494-900X; Search for: Sulea, Traian¹ORCID identifier: https://orcid.org/0000-0001-5301-8261
Affiliation	National Research Council of Canada. Human Health Therapeutics
Format	Text, Article
Subject	aggregation; native folding; single point mutation; structure-aggregation relationship; prediction method
Abstract	Solution stability is an important factor in the optimization of engineered biotherapeutic candidates such as monoclonal antibodies because of its possible effects on manufacturability, pharmacology, efficacy and safety. A detailed atomic understanding of the mechanisms governing self-association of natively folded protein monomers is required to devise predictive tools to guide screening and re-engineering along the drug development pipeline. We investigated pairs of affinity-matured full-size antibodies and observed drastically different propensities to aggregate from variants differing by a single amino-acid. Biophysical testing showed that antigen-binding fragments (Fabs) from the aggregating antibodies also reversibly associated with equilibrium dissociation constants in the low-micromolar range. Crystal structures (PDB accession codes 6MXR, 6MXS, 6MY4, 6MY5) and bottom-up hydrogen-exchange mass spectrometry revealed that Fab self-association occurs in a symmetric mode that involves the antigen complementarity-determining regions. Subtle local conformational changes incurred upon point mutation of monomeric variants foster formation of complementary polar interactions and hydrophobic contacts to generate a dimeric Fab interface. Testing of popular in silico tools generally indicated low reliabilities for predicting the aggregation propensities observed. A structure-aggregation data set is provided here in order to stimulate further improvements of in silico tools for prediction of native aggregation. Incorporation of intermolecular docking, conformational flexibility, and short-range packing interactions may all be necessary features of the ideal algorithm.
Publication date	2019-07-19
Publisher	Taylor & Francis
Licence	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) https://creativecommons.org/licenses/by-nc-nd/4.0/
In	mAbs 11, no. 7: 1300–1318.
Language	English
Peer reviewed	Yes
NRC number	NRC-HHT_53508
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Record identifier	0c949240-61b0-4677-8818-225315b4f12a
Record created	2020-11-27
Record modified	2021-01-08

Date modified:: 2024-07-11