Novel multivalent design of a monoclonal antibody improves binding strength to soluble aggregates of amyloid beta  

作　　者：Fadi Rofo Jos Buijs Ronny Falk Ken Honek Lars Lannfelt Anna M.Lilja Nicole G.Metzendorf Tobias Gustavsson Dag Sehlin Linda Söderberg Greta Hultqvist 

机构地区：[1]Protein Drug Design,Faculty of Pharmacy,Uppsala University,75124 Uppsala,Sweden [2]Department of Immunology,Genetics and Pathology,Uppsala University,75185 Uppsala,Sweden [3]Ridgeview Instruments,75237 Uppsala,Sweden [4]BioArctic AB,11251 Stockholm,Sweden [5]Department of Public Health and Caring Sciences,Uppsala University,75185 Uppsala,Sweden

出　　处：《Translational Neurodegeneration》2021年第3期502-517,共16页转化神经变性病（英文）

基　　金：This work was supported by grants from Swedish Research Council,Hedlunds stiftelse,Åke Wibergs stiftelse,Åhlen-stiftelsen,Jeanssons stiftelser,Magnus Bergvalls stiftelse,Vinnova and Alzheimerfonden。

摘　　要：Background:Amyloid-β(Aβ)immunotherapy is a promising therapeutic strategy in the fght 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 efcacy or side-efects,two antibodies are currently in phase 3,and one has been approved by FDA.The soluble intermediate aggregated species of Aβ,termed oligomers and protofbrils,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 Aβprotofbrilselective 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 Aβprotofbrils.Diferent ELISA setups were applied to demonstrate the binding strength of the hexavalent antibody to Aβaggregates of diferent sizes.Finally,the ability of the antibodies to protect cells from Aβ-induced efects 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 protofbrils,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 fbrils.The hexavalent antibody also reduced cell death induced by a mixture of soluble Aβaggregates.Conclusion:We provide a new antibody design with increased valency to promote binding av

关 键 词：Multivalent antibodies Alzheimer’s disease Aβ AVIDITY OLIGOMERS Protofbrils 

分 类 号：R749.16[医药卫生—神经病学与精神病学]