| Literature DB >> 35798951 |
M Rodrigues1,2, P Bhattacharjee3,4, A Brinkmalm3,4, D T Do5, C M Pearson5, S De1,2,6, A Ponjavic1,7,8, J A Varela1,9, K Kulenkampff1,2, I Baudrexel1, D Emin1,2, F S Ruggeri1,10,11, J E Lee1,12, A R Carr1, T P J Knowles1, H Zetterberg3,4,13,14, T N Snaddon15, S Gandhi16,17, S F Lee18, D Klenerman19,20.
Abstract
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.Entities:
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Year: 2022 PMID： 35798951 DOI： 10.1038/s41557-022-00976-3
Source DB: PubMed Journal: Nat Chem ISSN： 1755-4330 Impact factor: 24.274