Binding symmetry and surface flexibility mediate antibody self-association
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...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2019-10-01
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| Series: | mAbs |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2019.1632114 |
| _version_ | 1811342677002682368 |
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| author | Joseph D. Schrag Marie-Ève Picard Francis Gaudreault Louis-Patrick Gagnon Jason Baardsnes Mahder S. Manenda Joey Sheff Christophe Deprez Cassio Baptista Hervé Hogues John F. Kelly Enrico O. Purisima Rong Shi Traian Sulea |
| author_facet | Joseph D. Schrag Marie-Ève Picard Francis Gaudreault Louis-Patrick Gagnon Jason Baardsnes Mahder S. Manenda Joey Sheff Christophe Deprez Cassio Baptista Hervé Hogues John F. Kelly Enrico O. Purisima Rong Shi Traian Sulea |
| author_sort | Joseph D. Schrag |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T19:15:04Z |
| format | Article |
| id | doaj.art-ce79077ec94f4690a1f6d8fff8b9138a |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2024-04-13T19:15:04Z |
| publishDate | 2019-10-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-ce79077ec94f4690a1f6d8fff8b9138a2022-12-22T02:33:42ZengTaylor & Francis GroupmAbs1942-08621942-08702019-10-011171300131810.1080/19420862.2019.1632114Binding symmetry and surface flexibility mediate antibody self-associationJoseph D. Schrag0Marie-Ève Picard1Francis Gaudreault2Louis-Patrick Gagnon3Jason Baardsnes4Mahder S. Manenda5Joey Sheff6Christophe Deprez7Cassio Baptista8Hervé Hogues9John F. Kelly10Enrico O. Purisima11Rong Shi12Traian Sulea13Human Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaDépartement de Biochimie, de Microbiologie et de Bio-informatique, PROTEO, and Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Pavillon Charles-Eugène-Marchand, Québec City, QC G1V 0A6, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaDépartement de Biochimie, de Microbiologie et de Bio-informatique, PROTEO, and Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Pavillon Charles-Eugène-Marchand, Québec City, QC G1V 0A6, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON K1A 0R6, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON K1A 0R6, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaDépartement de Biochimie, de Microbiologie et de Bio-informatique, PROTEO, and Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Pavillon Charles-Eugène-Marchand, Québec City, QC G1V 0A6, CanadaHuman Health Therapeutics Research Centre, National Research Council Canada, Montreal, QC H4P 2R2, CanadaSolution 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.https://www.tandfonline.com/doi/10.1080/19420862.2019.1632114Aggregationnative foldingsingle point mutationstructure-aggregation relationshipprediction method |
| spellingShingle | Joseph D. Schrag Marie-Ève Picard Francis Gaudreault Louis-Patrick Gagnon Jason Baardsnes Mahder S. Manenda Joey Sheff Christophe Deprez Cassio Baptista Hervé Hogues John F. Kelly Enrico O. Purisima Rong Shi Traian Sulea Binding symmetry and surface flexibility mediate antibody self-association mAbs Aggregation native folding single point mutation structure-aggregation relationship prediction method |
| title | Binding symmetry and surface flexibility mediate antibody self-association |
| title_full | Binding symmetry and surface flexibility mediate antibody self-association |
| title_fullStr | Binding symmetry and surface flexibility mediate antibody self-association |
| title_full_unstemmed | Binding symmetry and surface flexibility mediate antibody self-association |
| title_short | Binding symmetry and surface flexibility mediate antibody self-association |
| title_sort | binding symmetry and surface flexibility mediate antibody self association |
| topic | Aggregation native folding single point mutation structure-aggregation relationship prediction method |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2019.1632114 |
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