Separation and detection of individual A尾 aggregates by capillary electrophoresis with laser-induced fluorescence detection

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• 作者：Ryan A. Picou^a ; David P. Schrum^b ; Ginger Ku^c ; Rebekah A. Cerqua^a ; Indu Kheterpal^c ; S. Douglass Gilman^a ; ^{sdgilman@lsu.edu}
• 关键词：Capillary electrophoresis ; Fluorescence ; Amyloid ; Fluorescence anisotropy ; Protein aggregation
• 刊名：Analytical Biochemistry
• 出版年：2012
• 期刊代码：93_00032697
• 类别：imm
• 出版时间：15 June, 2012
• 卷：425
• 期：2
• 页码：104-112
• 文件大小：873 K

摘要

The separation and detection of individual amyloid beta (A尾) aggregates by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was demonstrated. Samples were prepared with either A尾 (1-40) or A尾 (1-42) peptides and were characterized by CE with ultraviolet (UV) absorbance detection and transmission electron microscopy (TEM). Using thioflavin T (ThT) in the electrophoresis buffer, electrophoresis of aggregate-containing samples (5.0-s injection) produced up to several hundred narrow (<20 ms FWHM [full width at half maximum]) fluorescence peaks. Injection of A尾 (1-40) monomer samples resulted in no additional peaks compared with controls. The CE-LIF results were validated by bulk ThT fluorescence measurements for the same samples. The potential of laser-induced fluorescence anisotropy (LIFA) with CE to characterize individual A尾 aggregates also was investigated.