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

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

doi:10.1186/s40035-021-00258-x

Cited by 32 publications

(55 citation statements)

References 60 publications

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“…We have previously introduced HexaRmA158 (Fig. 1c), a multivalent antibody with extra binding sites in the form of scFv, providing a 40 times enhanced avidity and slower rate of dissociation from Aβ protofibrils compared to RmAb158 [23]. Importantly, due to the short distance between the additional binding sites, the multivalent antibody could bind strongly and with avidity to small Aβ oligomers [23], an effect that has been difficult to achieve with conventional IgG antibodies [20][21][22].…”

Section: Discussionmentioning

confidence: 99%

“…1c), a multivalent antibody with extra binding sites in the form of scFv, providing a 40 times enhanced avidity and slower rate of dissociation from Aβ protofibrils compared to RmAb158 [23]. Importantly, due to the short distance between the additional binding sites, the multivalent antibody could bind strongly and with avidity to small Aβ oligomers [23], an effect that has been difficult to achieve with conventional IgG antibodies [20][21][22]. HexaRmAb158 reduced neuronal cell death in vitro when added to cell cultures together with soluble Aβ aggregates [23], suggesting the potential of this antibody format to generate future therapeutic interventions targeting toxic Aβ aggregates in vivo.…”

Section: Discussionmentioning

confidence: 99%

“…In the current study, both antibody formats were essentially equal in lowering the concentration of soluble Aβ aggregates in vivo. The multivalent antibody format (Hex-aRmAb158) has previously demonstrated a stronger binding than RmAb158 to Aβ oligomers in vitro [23]. These species of Aβ should be highly present in the TBS soluble brain extracts ultra-centrifuged at 100,000 × g and 200,000 × g. However, these fractions might also contain soluble Aβ aggregates other than oligomers, such as the bigger protofibrils [60] or even smaller aggregates like trimers/dimers [61].…”

Section: Discussionmentioning

confidence: 99%

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A Brain-Targeting Bispecific-Multivalent Antibody Clears Soluble Amyloid-Beta Aggregates in Alzheimer's Disease Mice

et al. 2022

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Amyloid-β (Aβ) oligomers and protofibrils are suggested to be the most neurotoxic Aβ species in Alzheimer’s disease (AD). Hence, antibodies with strong and selective binding to these soluble Aβ aggregates are of therapeutic potential. We have recently introduced HexaRmAb158, a multivalent antibody with additional Aβ-binding sites in the form of single-chain fragment variables (scFv) on the N-terminal ends of Aβ protofibril selective antibody (RmAb158). Due to the additional binding sites and the short distance between them, HexaRmAb158 displayed a slow dissociation from protofibrils and strong binding to oligomers in vitro. In the current study, we aimed at investigating the therapeutic potential of this antibody format in vivo using mouse models of AD. To enhance BBB delivery, the transferrin receptor (TfR) binding moiety (scFv8D3) was added, forming the bispecific-multivalent antibody (HexaRmAb158-scFv8D3). The new antibody displayed a weaker TfR binding compared to the previously developed RmAb158-scFv8D3 and was less efficiently transcytosed in a cell-based BBB model. HexaRmAb158 detected soluble Aβ aggregates derived from brains of tg-ArcSwe and AppNL−G−F mice more efficiently compared to RmAb158. When intravenously injected, HexaRmAb158-scFv8D3 was actively transported over the BBB into the brain in vivo. Brain uptake was marginally lower than that of RmAb158-scFv8D3, but significantly higher than observed for conventional IgG antibodies. Both antibody formats displayed similar brain retention (72 h post injection) and equal capacity in clearing soluble Aβ aggregates in tg-ArcSwe mice. In conclusion, we demonstrate a bispecific-multivalent antibody format capable of passing the BBB and targeting a wide-range of sizes of soluble Aβ aggregates.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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A Brain-Targeting Bispecific-Multivalent Antibody Clears Soluble Amyloid-Beta Aggregates in Alzheimer's Disease Mice

et al. 2022

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“…Clazakizumab is beneficial not only for antibody-mediated rejection ( 51 ), but for active psoriatic arthritis ( 52 ). BAN2401 is mainly utilized for Alzheimer’s disease due to its advantage of improving binding strength to soluble aggregates of amyloid-beta ( 53 , 54 ). Although these drugs have not yet been currently used in SCI-induced OP, the results of text mining and computational analysis demonstrated that hub genes are regulated after SCI involved in the OP occurrence, suggesting that drugs targetable key gene symbols have the potential to prevent the occurrence and development of SCI-induced OP.…”

Section: Discussionmentioning

confidence: 99%

Drug discovery in spinal cord injury-induced osteoporosis: a text mining-based study

Wang¹,

Xu²,

Han³

et al. 2022

Ann Transl Med

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Background Spinal cord injury (SCI) and osteoporosis (OP) are common diseases in spine surgery, and OP could be the complication of SCI. However, SCI-induced OP is a complex pathologic process and drug discovery is limited, which restricts the study in the mechanism and treatment of the disease. This study aims to identify the genes and molecular pathways related to SCI-induced OP through computational tools and public datasets, and to explore drug targeting therapy, ultimately preventing the occurrence of OP after SCI. Methods In this study, common genes related to SCI and OP were obtained by text mining, then which conducted the functional analysis. Protein-protein interaction (PPI) networks were constructed by STRING online and Cytoscape software. Finally, core genes and potential drugs were performed after undergoing drug-gene interaction analysis which also completed functional analysis. Results A total of 371 genes common to ‘SCI’ and ‘OP’ were identified by text mining. After functional analysis, 207 significant genes were screened out. Subsequently, PPI analysis yielded 23 genes targetable by 13 drugs which were the candidate to treat SCI-induced OP. Conclusions Taken together, siltuximab, olokizumab, clazakizumab and BAN2401 were first discovered to become the potential drugs for the treatment of SCI-induced OP. Drug discovery using text mining and pathway analysis is a significant way to investigate the pathomechanism of the disease while exploring existing drugs to treat the disease.

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“…2A). Structural stability of the proteins was evaluated under thermal stress as described previously [40]. sNEP yielded an in ection temperature of 58 °C (Fig.…”

Section: In Vitro Characterization Of the Recombinant Proteinsmentioning

confidence: 99%

Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease

Rofo

Metzendorf

Saubi

et al. 2022

Preprint

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BackgroundAggregation of the amyloid-b (Ab) peptide in the brain is one of the key pathological events in Alzheimer’s disease (AD). Reducing Ab levels in the brain by enhancing its degradation is one of the important strategies to develop new therapies for AD. Neprilysin (NEP) is a membrane bound metallopeptidase and one of the major Ab-degrading enzymes. The secreted soluble form of NEP (sNEP) has been previously suggested as a potential protein-therapy degrading Ab in AD. However, similar to other large molecule, peripherally administered sNEP is unable to reach the brain due to the presence of the blood-brain barrier (BBB). Methods To provide transcytosis across BBB, we recombinantly fused the TfR binding moiety (scFv8D3) to either sNEP or to a previously described variant of NEP (muNEP) having higher degradation efficiency on Ab compared to other NEP substrates. To provide long blood half-life, an Fc-based antibody fragment (scFc) was added to the designs, forming sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3. Ability of the mentioned recombinant proteins to degrade Ab was first evaluated in vitro using synthetic Ab peptides followed by sandwich ELISA. For the in vivo studies, a single injection of 125-iodine labelled sNEP-scFc-scFv8D3 and muNEP-scFc-scFv8D3 was intravenously administered to tg-ArcSwe mouse model of AD, using scFc-scFv8D3 protein that lacks NEP as a negative control. Different ELISA setups were applied to quantify Ab concentration of different conformations, both in brain tissues and blood samples. ResultsWhen tested in vitro, sNEP-scFc-scFv8D3 retained sNEP enzymatic activity in degrading Ab. When intravenously injected, sNEP-scFc-scFv8D3 demonstrated 20-times higher brain uptake compared to sNEP. Both scFv8D3 fused NEP-proteins significantly reduced Ab levels in the blood of tg-ArcSwe mice, a transgenic mouse model of AD, following a single intravenous injection. However, the degradation of brain Ab was only evident with sNEP-scFc-scFv8D3 treatment, and both scFv8D3 fused NEP-proteins displayed a fast clearance from the brain. Conclusion Our study highlights the significance of BBB-penetrant proteins to study in details the effects of NEP on AD brains, avoiding the use of invasive intracranial administration or generation of NEP transgenic mice.

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

Cited by 32 publications

References 60 publications

A Brain-Targeting Bispecific-Multivalent Antibody Clears Soluble Amyloid-Beta Aggregates in Alzheimer's Disease Mice

A Brain-Targeting Bispecific-Multivalent Antibody Clears Soluble Amyloid-Beta Aggregates in Alzheimer's Disease Mice

Drug discovery in spinal cord injury-induced osteoporosis: a text mining-based study

Blood-brain barrier penetrating neprilysin degrades monomeric amyloid-beta in a mouse model of Alzheimer’s disease

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