Structure-specific amyloid precipitation in biofluids
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The composition of soluble toxic protein aggregates formed in vivo is currently unknown in neurodegenerative diseases, due to their ultra-low concentration in human biofluids and their high degree of heterogeneity. Here we report a method to capture amyloid-containing aggregates in human biofluids in an unbiased way, a process we name amyloid precipitation. We use a structure-specific chemical dimer, a Y-shaped, bio-inspired small molecule with two capture groups, for amyloid precipitation to increase affinity. Our capture molecule for amyloid precipitation (CAP-1) consists of a derivative of Pittsburgh Compound B (dimer) to target the cross β-sheets of amyloids and a biotin moiety for surface immobilization. By coupling CAP-1 to magnetic beads, we demonstrate that we can target the amyloid structure of all protein aggregates present in human cerebrospinal fluid, isolate them for analysis and then characterize them using single-molecule fluorescence imaging and mass spectrometry. Amyloid precipitation enables unbiased determination of the molecular composition and structural features of the in vivo aggregates formed in neurodegenerative diseases.
Rodrigues , M , Bhattacharjee , P , Brinkmalm , A , Do , D T , Pearson , C M , De , S , Ponjavic , A , Varela , J A , Kulenkampff , K , Baudrexel , I , Emin , D , Ruggeri , F S , Lee , J E , Carr , A R , Knowles , T P J , Zetterberg , H , Snaddon , T N , Gandhi , S , Lee , S F & Klenerman , D 2022 , ' Structure-specific amyloid precipitation in biofluids ' , Nature Chemistry , vol. 14 , no. 9 , pp. 1045-1056 . https://doi.org/10.1038/s41557-022-00976-3
Copyright © 2022, The Author(s), under exclusive licence to Springer Nature Limited. This work has been made available online in accordance with publisher policies or with permission. Permission for further reuse of this content should be sought from the publisher or the rights holder. This is the author created accepted manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at https://doi.org/10.1038/s41557-022-00976-3.
DescriptionFunding: This work was supported by the National Institute of Health Research University College London Hospitals Biomedical Research Centre. SG is an MRC Senior Clinical Fellow (MR/T008199/1). H.Z. is a Wallenberg Scholar supported by grants from the Swedish Research Council (no. 2018-02532), the European Research Council (no. 681712), Swedish State Support for Clinical Research (no. ALFGBG-720931) and the UK Dementia Research Institute at University College London (UCL). D.K. is supported by grants from the European Research Council (no. 669237), the Royal Society and the UK Dementia Research Institute at Cambridge. We thank the Royal Society for the University Research Fellowship to S.F.L. (UF120277), and T.N.S. thanks the National Institutes of Health (R01GM121573). Also, we are thankful for the Michael J. Fox Grant to S.F.L. and T.N.S. (grant no. 10200). J.A.V. is supported by the European Research Council with an ERC Starting Grant (no. 804581).
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