Binding symmetry and surface flexibility mediate antibody self-association.
Antibodies, Monoclonal
/ chemistry
Bioengineering
Complementarity Determining Regions
/ chemistry
Dimerization
Humans
Immunoglobulin Fab Fragments
/ chemistry
Mass Spectrometry
Mutation
/ genetics
Protein Aggregates
Protein Conformation
Protein Folding
Protein Stability
Stereoisomerism
Structure-Activity Relationship
Aggregation
native folding
prediction method
single point mutation
structure-aggregation relationship
Journal
mAbs
ISSN: 1942-0870
Titre abrégé: MAbs
Pays: United States
ID NLM: 101479829
Informations de publication
Date de publication:
10 2019
10 2019
Historique:
pubmed:
19
7
2019
medline:
18
7
2020
entrez:
19
7
2019
Statut:
ppublish
Résumé
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
Identifiants
pubmed: 31318308
doi: 10.1080/19420862.2019.1632114
pmc: PMC6748613
doi:
Substances chimiques
Antibodies, Monoclonal
0
Complementarity Determining Regions
0
Immunoglobulin Fab Fragments
0
Protein Aggregates
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1300-1318Références
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