Structure-specific amyloid precipitation in biofluids

M. Rodrigues; P. Bhattacharjee; A. Brinkmalm; D.T. Do; C.M. Pearson; S. De; A. Ponjavic; J.A. Varela; K. Kulenkampff; I. Baudrexel; D. Emin; F.S. Ruggeri; J.E. Lee; A.R. Carr; T.P.J. Knowles; H. Zetterberg; T.N. Snaddon; S. Gandhi; S.F. Lee; D. Klenerman

doi:10.1038/s41557-022-00976-3

Structure-specific amyloid precipitation in biofluids

M. Rodrigues, P. Bhattacharjee, A. Brinkmalm, D.T. Do, C.M. Pearson, S. De, A. Ponjavic, J.A. Varela, K. Kulenkampff, I. Baudrexel, D. Emin, F.S. Ruggeri, J.E. Lee, A.R. Carr, T.P.J. Knowles, H. Zetterberg, T.N. Snaddon^*, S. Gandhi^*, S.F. Lee^*, D. Klenerman^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	1045-1056
Number of pages	12
Journal	Nature Chemistry
Volume	14
Issue number	9
Early online date	7 Jul 2022
DOIs	https://doi.org/10.1038/s41557-022-00976-3
Publication status	Published - Sept 2022

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1038/s41557-022-00976-3

Rodrigues-2022-Structure-specific-NatChem-AAM
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.
Accepted author manuscript, 10 MB

https://www.repository.cam.ac.uk/handle/1810/337289

BrainNanoFlow: Nanoscale dynamics: H2020 ERC Starting Grant 2018 BrainNanoFlow Juan Varela
Varela, J. A. (PI)
European Research Council
1/12/18 → 30/11/23
Project: Fellowship

Cite this

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, 14(9), 1045-1056. https://doi.org/10.1038/s41557-022-00976-3

@article{c629169bf1db4bdeb033f1270e74d8c4,

title = "Structure-specific amyloid precipitation in biofluids",

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.",

author = "M. Rodrigues and P. Bhattacharjee and A. Brinkmalm and D.T. Do and C.M. Pearson and S. De and A. Ponjavic and J.A. Varela and K. Kulenkampff and I. Baudrexel and D. Emin and F.S. Ruggeri and J.E. Lee and A.R. Carr and T.P.J. Knowles and H. Zetterberg and T.N. Snaddon and S. Gandhi and S.F. Lee and D. Klenerman",

note = "Funding: 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).",

year = "2022",

month = sep,

doi = "10.1038/s41557-022-00976-3",

language = "English",

volume = "14",

pages = "1045--1056",

journal = "Nature Chemistry",

issn = "1755-4330",

publisher = "Nature publishing group",

number = "9",

}

Rodrigues, M, Bhattacharjee, P, Brinkmalm, A, Do, DT, Pearson, CM, De, S, Ponjavic, A, Varela, JA, Kulenkampff, K, Baudrexel, I, Emin, D, Ruggeri, FS, Lee, JE, Carr, AR, Knowles, TPJ, Zetterberg, H, Snaddon, TN, Gandhi, S, Lee, SF & 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

TY - JOUR

T1 - Structure-specific amyloid precipitation in biofluids

AU - Rodrigues, M.

AU - Bhattacharjee, P.

AU - Brinkmalm, A.

AU - Do, D.T.

AU - Pearson, C.M.

AU - De, S.

AU - Ponjavic, A.

AU - Varela, J.A.

AU - Kulenkampff, K.

AU - Baudrexel, I.

AU - Emin, D.

AU - Ruggeri, F.S.

AU - Lee, J.E.

AU - Carr, A.R.

AU - Knowles, T.P.J.

AU - Zetterberg, H.

AU - Snaddon, T.N.

AU - Gandhi, S.

AU - Lee, S.F.

AU - Klenerman, D.

N1 - Funding: 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).

PY - 2022/9

Y1 - 2022/9

N2 - 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.

AB - 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.

U2 - 10.1038/s41557-022-00976-3

DO - 10.1038/s41557-022-00976-3

M3 - Article

SN - 1755-4330

VL - 14

SP - 1045

EP - 1056

JO - Nature Chemistry

JF - Nature Chemistry

IS - 9

ER -

Structure-specific amyloid precipitation in biofluids

Abstract

UN SDGs

Access to Document

Fingerprint

Projects

BrainNanoFlow: Nanoscale dynamics: H2020 ERC Starting Grant 2018 BrainNanoFlow Juan Varela

Cite this