Reproducible Formation of Insulin Superstructures: Amyloid-Like Fibrils, Spherulites, and Particulates
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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Reproducible Formation of Insulin Superstructures : Amyloid-Like Fibrils, Spherulites, and Particulates. / Thorlaksen, Camilla; Neergaard, Martin Busch; Groenning, Minna; Foderà, Vito.
Protein Aggregation: Methods in Molecular Biology. ed. / A. Stanisław Cieplak. Humana Press, 2023. p. 297-309 (Methods in Molecular Biology, Vol. 2551).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Reproducible Formation of Insulin Superstructures
T2 - Amyloid-Like Fibrils, Spherulites, and Particulates
AU - Thorlaksen, Camilla
AU - Neergaard, Martin Busch
AU - Groenning, Minna
AU - Foderà, Vito
N1 - Funding Information: The work was funded, and all protein material was provided by Novo Nordisk A/S. For use of the TEM, we acknowledge the Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen. For use of the MFI image evaluation software, we acknowledge Jesper Søndergaard Marino from Novo Nordisk A/S. Illustrations presented in the chapter were created with Biorender.com. V.F. also acknowledges the VIL-LUM FONDEN for the Villum Young Investigator Grant “Protein Superstructures as Smart Biomaterials (ProSmart)” 2018–2023 (project number: 19175). The authors acknowledge Marco van de Weert (University of Copenhagen) for inspiring discussions on reproducibility in protein stability and aggregation studies. Publisher Copyright: © 2023, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2023
Y1 - 2023
N2 - Inducing protein aggregation in vitro under various formulation and stress conditions may lead to an increased understanding of the different association routes a protein can undergo. However, a range of factors can affect the aggregation process, often leading to heterogenous samples and experimental irreproducibility between labs. Here, we present detailed methods to reproducibly form homogenous samples of superstructures: amyloid-like fibrils, spherulites, and particulates from human insulin. We discuss pitfalls and good practice in the lab, with the aim of creating awareness on the potential sources of artefacts for protein stability and aggregation studies.
AB - Inducing protein aggregation in vitro under various formulation and stress conditions may lead to an increased understanding of the different association routes a protein can undergo. However, a range of factors can affect the aggregation process, often leading to heterogenous samples and experimental irreproducibility between labs. Here, we present detailed methods to reproducibly form homogenous samples of superstructures: amyloid-like fibrils, spherulites, and particulates from human insulin. We discuss pitfalls and good practice in the lab, with the aim of creating awareness on the potential sources of artefacts for protein stability and aggregation studies.
KW - Aggregation
KW - Amyloid-like fibrils
KW - Insulin
KW - Micro-Flow Imaging
KW - Particulates
KW - Spherulites
KW - Superstructures
KW - Transmission electron microscopy
U2 - 10.1007/978-1-0716-2597-2_20
DO - 10.1007/978-1-0716-2597-2_20
M3 - Book chapter
C2 - 36310211
AN - SCOPUS:85141005126
SN - 978-1-0716-2596-5
T3 - Methods in Molecular Biology
SP - 297
EP - 309
BT - Protein Aggregation
A2 - Stanisław Cieplak, A.
PB - Humana Press
ER -
ID: 333701451