A modified protocol to prepare seed-free starting solutions of amyloid-β (Aβ)_1–40 and Aβ_1–42 from the corresponding depsipeptides

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• 作者：Marten Beeg¹ ; ² ; ^a ; ^{marten.beeg@chem.ethz.ch} ; Matteo Stravalaci² ; ^a ; Antonio Bastone^a ; Mario Salmona^a ; Marco Gobbi^a
• 刊名：Analytical Biochemistry
• 出版年：2011
• 出版时间：15 April 2011
• 年：2011
• 卷：411
• 期：2
• 页码：297-299
• 全文大小：356 K

文摘

Preparing reliable, seed-free stock solutions of the highly amyloidogenic peptides amyloid-β (Aβ) is difficult. Besides the formation of aggregates during synthesis and storage, dissolution of the peptide is a critical step because vortexing can induce aggregation. To overcome this, synthesis of the more water-soluble depsi-Aβ_1–42 peptide, from which the native sequence is easily obtained, has been suggested. We further refined this technique, including a cutoff filtration step and switching the depsipeptide in basic conditions, to stabilize the formed native peptide. The obtained solutions of native Aβ_1–40 and Aβ_1–42 peptides were homogeneous and aggregate free, as indicated by thioflavin T and circular dichroism analysis.