Tau Fibrillation Induced by Heparin or a Lysophospholipid Show Different Initial Oligomer Formation

Rasmussen, H.H.; Nielsen, Janni; Poli, Angela de; Otzen, Daniel E.; Pedersen, Jan Skov

doi:10.1016/j.jmb.2023.168194

Cited by 3 publications

(1 citation statement)

References 87 publications

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“…Absorption was finally measured at 450 and 620 nm on a HiPo Microplate Photometer MPP-96 (BioSan, Riga, Latvia). To test for binding to Aβ40 and tau, we used 2 µg/mL Aβ40 (Chinese Peptide, 95% purity) and 2 µg/ml tau produced recombinantly as described 53 and incubated with a dilution series of 5 different mAbs (10-9C8-C1, 14-9E7-A1, 20-9E2-B8, 17 9D12 A1 and 18 3A5H2) from 10 µg/ml and down in 10 steps of 2-fold dilution, followed by the steps described above. As positive controls we used anti-tau antibodies 46 32274, 5 58860 and 13 21796 (Santa Cruz, CA).…”

Section: Indirect Elisa Assaysmentioning

confidence: 99%

Monoclonal antibodies targeting cytotoxic α-synuclein oligomers: molecular properties and diagnostic potential

Otzen,

Nielsen,

Lauritsen

et al. 2023

Preprint

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Aggregation of α-synuclein (α-syn) is critical to the development of synucleinopathies such as Parkinson’s Disease (PD), yet we do not have any effective therapy against these diseases. α-syn accumulates as insoluble amyloid in intracellular Lewy Bodies (LBs) but also forms soluble αSOs (αSOs) which are thought to be even more cytotoxic than fibrils. We lack tools to detect and block the unwanted activities of these αSOs. To address this, we have raised monoclonal antibodies (mAbs) against different forms of αSOs, ranging from unmodified αSOs to species stabilized by lipid peroxidation products and polyphenols, αSOs formed by C-terminally truncated α-syn, and multivalent display of α-syn on capsid virus-like particles (cVLPs). Here we report on the general biophysical and immunohistochemical properties of 30 anti-αSO mAbs obtained in this fashion. Our analysis includes direct measurement of in-solution affinity to monomeric α-syn and αSOs using Flow-Induced Dispersion Analysis, which avoids immobilization artifacts associated with ELISA. While the mAbs generally show a preference for αSOs, they also bind fibrils, but to variable extents. Overall, we observe great diversity in the mAbs’ relative affinities for monomers and αSOs, varied requirements for the C-terminal extension of α-syn, and only a modest effect on α-syn fibrillation. Several mAbs show several orders of magnitude preference for αSOs over monomers in in-solution studies, in contrast to the commercial antibody MJF14 which only bound 10-fold more strongly to αSOs than monomeric α-syn. Gratifyingly, seven mAbs almost completely block αSO permeabilization of membrane vesicles. 16 mAbs were analyzed for binding to brain sections from rats overexpressing human α-syn in the nigro-striatal pathway, and the five most promising were further investigated for their ability to recognize α-syn aggregates in human brain tissue. All five mAbs identified α-syn-related pathologies like LBs and Lewy Neurites, as well as Glial Cytoplasmic Inclusions in postmortem brains from people diagnosed for PD, dementia with LBs and multiple system atrophy, although to different extents. We have identified three mAbs as particularly useful for pathological evaluation of postmortem brain human tissue, including the early stages of PD. Although there was no straightforward connection between the individual mAbs’ biophysical and immunohistochemical properties, it is encouraging that a large collection of mAbs able to recognize different aggregated α-syn species in vitro also hold diagnostic potential.

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Section: Indirect Elisa Assaysmentioning

confidence: 99%

Monoclonal antibodies targeting cytotoxic α-synuclein oligomers: molecular properties and diagnostic potential

Otzen,

Nielsen,

Lauritsen

et al. 2023

Preprint

Self Cite

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Molecular properties and diagnostic potential of monoclonal antibodies targeting cytotoxic α-synuclein oligomers

Nielsen,

Lauritsen,

Pedersen

et al. 2024

npj Parkinsons Dis.

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Abstractα-Synuclein (α-syn) accumulates as insoluble amyloid but also forms soluble α-syn oligomers (αSOs), thought to be even more cytotoxic than fibrils. To detect and block the unwanted activities of these αSOs, we have raised 30 monoclonal antibodies (mAbs) against different forms of αSOs, ranging from unmodified αSOs to species stabilized by lipid peroxidation products and polyphenols, αSOs formed by C-terminally truncated α-syn, and multivalent display of α-syn on capsid virus-like particles (cVLPs). While the mAbs generally show a preference for αSOs, they also bind fibrils, but to variable extents. Overall, we observe great diversity in the mAbs’ relative affinities for monomers and αSOs, varied requirements for the C-terminal extension of α-syn, and only a modest effect on α-syn fibrillation. Several mAbs show several orders of magnitude preference for αSOs over monomers in in-solution studies, while the commercial antibody MJF14 only bound 10-fold more strongly to αSOs than monomeric α-syn. Gratifyingly, seven mAbs almost completely block αSO permeabilization of membrane vesicles. Five selected mAbs identified α-syn-related pathologies like Lewy bodies (LBs) and Lewy Neurites, as well as Glial Cytoplasmic Inclusions in postmortem brains from people diagnosed for PD, dementia with LBs or multiple system atrophy, although to different extents. Three mAbs were particularly useful for pathological evaluation of postmortem brain human tissue, including early stages of PD. Although there was no straightforward connection between the mAbs’ biophysical and immunohistochemical properties, it is encouraging that this comprehensive collection of mAbs able to recognize different aggregated α-syn species in vitro also holds diagnostic potential.

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Structures of Oligomeric States of Tau Protein, Amyloid-β, α-Synuclein and Prion Protein Implicated in Alzheimer’s Disease, Parkinson’s Disease and Prionopathies

Cehlar,

Njemoga,

Horvath

et al. 2024

IJMS

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In this review, we focus on the biophysical and structural aspects of the oligomeric states of physiologically intrinsically disordered proteins and peptides tau, amyloid-β and α-synuclein and partly disordered prion protein and their isolations from animal models and human brains. These protein states may be the most toxic agents in the pathogenesis of Alzheimer’s and Parkinson’s disease. It was shown that oligomers are important players in the aggregation cascade of these proteins. The structural information about these structural states has been provided by methods such as solution and solid-state NMR, cryo-EM, crosslinking mass spectrometry, AFM, TEM, etc., as well as from hybrid structural biology approaches combining experiments with computational modelling and simulations. The reliable structural models of these protein states may provide valuable information for future drug design and therapies.

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Tau Fibrillation Induced by Heparin or a Lysophospholipid Show Different Initial Oligomer Formation

Cited by 3 publications

References 87 publications

Monoclonal antibodies targeting cytotoxic α-synuclein oligomers: molecular properties and diagnostic potential

Monoclonal antibodies targeting cytotoxic α-synuclein oligomers: molecular properties and diagnostic potential

Molecular properties and diagnostic potential of monoclonal antibodies targeting cytotoxic α-synuclein oligomers

Structures of Oligomeric States of Tau Protein, Amyloid-β, α-Synuclein and Prion Protein Implicated in Alzheimer’s Disease, Parkinson’s Disease and Prionopathies

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