期刊
PROTEIN ENGINEERING DESIGN & SELECTION
卷 24, 期 9, 页码 743-750出版社
OXFORD UNIV PRESS
DOI: 10.1093/protein/gzr020
关键词
aggregation; amyloid beta peptide; biocompatibility; in vitro studies; solubilization
资金
- Alzheimer Research Trust (UK) [ART/PG2007/3]
- FWO [G.0903.09]
- Belgian Science Policy Office (BELSPO) [P6/43]
- Stichting voor Alzheimer Onderzoek (SAO) [09005]
- Agency for Innovation by Science and Technology Belgium [083052]
- EMBO [ALTF-512-2008]
- MRC [MC_U117533887, MC_U117584256] Funding Source: UKRI
- Medical Research Council [MC_U117533887, MC_U117584256] Funding Source: researchfish
We provide a validated and rapid protocol for the solubilization of amyloid -peptide (A). This procedure involves sequential solubilization using structure-breaking organic solvents hexafluoroisopropanol and DMSO followed by column purification. The low solubility and tendency of A to aggregate considerably impede the in vitro handling and biophysical or biological investigation of A, despite the interest in this peptide because of its implication in Alzheimers disease. The main advantage of the proposed protocol over others is that it results in standardized aggregate-free A peptide samples that are biocompatible for cell culture studies and yield reproducible aggregation kinetics and cytotoxicities. This three-step protocol also enables the co-solubilization of the longer A42 variant with A40 in ratios relevant to Alzheimers disease.
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