Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta
Abstract Background Amyloid-β (Aβ) immunotherapy is a promising therapeutic strategy in the fight against Alzheimer’s disease (AD). A number of monoclonal antibodies have entered clinical trials for AD. Some of them have failed due to the lack of efficacy or side-effects, two antibodies are currentl...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | Translational Neurodegeneration |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40035-021-00258-x |
| _version_ | 1818731164998828032 |
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| author | Fadi Rofo Jos Buijs Ronny Falk Ken Honek Lars Lannfelt Anna M. Lilja Nicole G. Metzendorf Tobias Gustavsson Dag Sehlin Linda Söderberg Greta Hultqvist |
| author_facet | Fadi Rofo Jos Buijs Ronny Falk Ken Honek Lars Lannfelt Anna M. Lilja Nicole G. Metzendorf Tobias Gustavsson Dag Sehlin Linda Söderberg Greta Hultqvist |
| author_sort | Fadi Rofo |
| collection | DOAJ |
| description | Abstract Background Amyloid-β (Aβ) immunotherapy is a promising therapeutic strategy in the fight against Alzheimer’s disease (AD). A number of monoclonal antibodies have entered clinical trials for AD. Some of them have failed due to the lack of efficacy or side-effects, two antibodies are currently in phase 3, and one has been approved by FDA. The soluble intermediate aggregated species of Aβ, termed oligomers and protofibrils, are believed to be key pathogenic forms, responsible for synaptic and neuronal degeneration in AD. Therefore, antibodies that can strongly and selectively bind to these soluble intermediate aggregates are of great diagnostic and therapeutic interest. Methods We designed and recombinantly produced a hexavalent antibody based on mAb158, an Aβ protofibril-selective antibody. The humanized version of mAb158, lecanemab (BAN2401), is currently in phase 3 clinical trials for the treatment of AD. The new designs involved recombinantly fusing single-chain fragment variables to the N-terminal ends of mAb158 antibody. Real-time interaction analysis with LigandTracer and surface plasmon resonance were used to evaluate the kinetic binding properties of the generated antibodies to Aβ protofibrils. Different ELISA setups were applied to demonstrate the binding strength of the hexavalent antibody to Aβ aggregates of different sizes. Finally, the ability of the antibodies to protect cells from Aβ-induced effects was evaluated by MTT assay. Results Using real-time interaction analysis with LigandTracer, the hexavalent design promoted a 40-times enhanced binding with avidity to protofibrils, and most of the added binding strength was attributed to the reduced rate of dissociation. Furthermore, ELISA experiments demonstrated that the hexavalent design also had strong binding to small oligomers, while retaining weak and intermediate binding to monomers and insoluble fibrils. The hexavalent antibody also reduced cell death induced by a mixture of soluble Aβ aggregates. Conclusion We provide a new antibody design with increased valency to promote binding avidity to an enhanced range of sizes of Aβ aggregates. This approach should be general and work for any aggregated protein or repetitive target. |
| first_indexed | 2024-12-17T23:13:20Z |
| format | Article |
| id | doaj.art-e3112d378ac74be8899dc991771ba21a |
| institution | Directory Open Access Journal |
| issn | 2047-9158 |
| language | English |
| last_indexed | 2024-12-17T23:13:20Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
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| series | Translational Neurodegeneration |
| spelling | doaj.art-e3112d378ac74be8899dc991771ba21a2022-12-21T21:29:04ZengBMCTranslational Neurodegeneration2047-91582021-09-0110111610.1186/s40035-021-00258-xNovel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid betaFadi Rofo0Jos Buijs1Ronny Falk2Ken Honek3Lars Lannfelt4Anna M. Lilja5Nicole G. Metzendorf6Tobias Gustavsson7Dag Sehlin8Linda Söderberg9Greta Hultqvist10Protein Drug Design, Faculty of Pharmacy, Uppsala UniversityDepartment of Immunology, Genetics and Pathology, Uppsala UniversityBioArctic ABBioArctic ABBioArctic ABBioArctic ABProtein Drug Design, Faculty of Pharmacy, Uppsala UniversityDepartment of Public Health and Caring Sciences, Uppsala UniversityDepartment of Public Health and Caring Sciences, Uppsala UniversityBioArctic ABProtein Drug Design, Faculty of Pharmacy, Uppsala UniversityAbstract Background Amyloid-β (Aβ) immunotherapy is a promising therapeutic strategy in the fight against Alzheimer’s disease (AD). A number of monoclonal antibodies have entered clinical trials for AD. Some of them have failed due to the lack of efficacy or side-effects, two antibodies are currently in phase 3, and one has been approved by FDA. The soluble intermediate aggregated species of Aβ, termed oligomers and protofibrils, are believed to be key pathogenic forms, responsible for synaptic and neuronal degeneration in AD. Therefore, antibodies that can strongly and selectively bind to these soluble intermediate aggregates are of great diagnostic and therapeutic interest. Methods We designed and recombinantly produced a hexavalent antibody based on mAb158, an Aβ protofibril-selective antibody. The humanized version of mAb158, lecanemab (BAN2401), is currently in phase 3 clinical trials for the treatment of AD. The new designs involved recombinantly fusing single-chain fragment variables to the N-terminal ends of mAb158 antibody. Real-time interaction analysis with LigandTracer and surface plasmon resonance were used to evaluate the kinetic binding properties of the generated antibodies to Aβ protofibrils. Different ELISA setups were applied to demonstrate the binding strength of the hexavalent antibody to Aβ aggregates of different sizes. Finally, the ability of the antibodies to protect cells from Aβ-induced effects was evaluated by MTT assay. Results Using real-time interaction analysis with LigandTracer, the hexavalent design promoted a 40-times enhanced binding with avidity to protofibrils, and most of the added binding strength was attributed to the reduced rate of dissociation. Furthermore, ELISA experiments demonstrated that the hexavalent design also had strong binding to small oligomers, while retaining weak and intermediate binding to monomers and insoluble fibrils. The hexavalent antibody also reduced cell death induced by a mixture of soluble Aβ aggregates. Conclusion We provide a new antibody design with increased valency to promote binding avidity to an enhanced range of sizes of Aβ aggregates. This approach should be general and work for any aggregated protein or repetitive target.https://doi.org/10.1186/s40035-021-00258-xMultivalent antibodiesAlzheimer’s diseaseAβAvidityOligomersProtofibrils |
| spellingShingle | Fadi Rofo Jos Buijs Ronny Falk Ken Honek Lars Lannfelt Anna M. Lilja Nicole G. Metzendorf Tobias Gustavsson Dag Sehlin Linda Söderberg Greta Hultqvist Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta Translational Neurodegeneration Multivalent antibodies Alzheimer’s disease Aβ Avidity Oligomers Protofibrils |
| title | Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| title_full | Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| title_fullStr | Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| title_full_unstemmed | Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| title_short | Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| title_sort | novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta |
| topic | Multivalent antibodies Alzheimer’s disease Aβ Avidity Oligomers Protofibrils |
| url | https://doi.org/10.1186/s40035-021-00258-x |
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