Real-Time Monitoring of Alzheimer鈥檚-Related Amyloid Aggregation via Probe Enhancement鈥揊luorescence Correlation Spectroscopy

详细信息    查看全文

• 作者：Yinghua Guan ; Kevin J. Cao ; Adam Cantlon ; Kristyna Elbel ; Emmanuel A. Theodorakis ; Dominic M. Walsh ; Jerry Yang ; Jagesh V. Shah
• 刊名：ACS Chemical Neuroscience
• 出版年：2015
• 出版时间：September 16, 2015
• 年：2015
• 卷：6
• 期：9
• 页码：1503-1508
• 全文大小：303K
• ISSN：1948-7193

文摘

This work describes the use of fluorescence correlation spectroscopy (FCS) and a novel amyloid-binding fluorescent probe, ARCAM 1, to monitor the aggregation of the Alzheimer鈥檚 disease-associated amyloid 尾-peptide (A尾). ARCAM 1 exhibits a large increase in fluorescence emission upon binding to A尾 assemblies, making it an excellent candidate for probe enhancement FCS (PE-FCS). ARCAM 1 binding does not change A尾 aggregation kinetics. It also exhibits greater dynamic range as a probe in reporting aggregate size by FCS in A尾, when compared to thioflavin T (ThT) or an A尾 peptide modified with a fluorophore. Using fluorescent burst analysis (via PE-FCS) to follow aggregation of A尾, we detected soluble aggregates at significantly earlier time points compared to typical bulk fluorescence measurements. Autocorrelation analysis revealed the size of these early A尾 assemblies. These results indicate that PE-FCS/ARCAM 1 based assays can detect and provide size characterization of small A尾 aggregation intermediates during the assembly process, which could enable monitoring and study of such aggregates that transiently accumulate in biofluids of patients with Alzheimer鈥檚 and other neurodegenerative diseases.