Microchip Electrophoresis Profiling of Aβ Peptides in the Cerebrospinal Fluid of Patients with Alzheimer’s Disease

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• 作者：Mohamad Reza Mohamadi ; Zuzana Svobodova ; Romain Verpillot ; Hermann Esselmann ; Jens Wiltfang ; Markus Otto ; Myriam Taverna ; Zuzana Bilkova ; Jean-Louis Viovy
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：September 15, 2010
• 年：2010
• 卷：82
• 期：18
• 页码：7611-7617
• 全文大小：224K
• 年卷期：v.82,no.18(September 15, 2010)
• ISSN：1520-6882

文摘

The preferential aggregation of Aβ1-42 in amyloid plaques is one of the major neuropathological events in Alzheimer’s disease. This is accompanied by a relative reduction of the concentration of Aβ1-42 in the cerebrospinal fluid (CSF) of patients developing the signs of Alzheimer’s disease. Here, we describe a microchip gel electrophoresis method in polydimethylsiloxane (PDMS) chip that enables rapid profiling of major Aβ peptides in cerebrospinal fluid. To control the electroosmotic flow (EOF) in the PDMS channel and also to reduce the adsorption of the peptides to the surface of the channel, a new double coating using poly(dimethylacrylamide-co-allyl glycidyl ether) (PDMA-AGE) and methylcellulose−Tween-20 was developed. With this method, separation of five synthetic Aβ peptides (Aβ1-37, Aβ1-38, Aβ1-39, Aβ1-40, and Aβ1-42) was achieved, and relative abundance of Aβ1-42 to Aβ1-37 could be calculated in different standard mixtures. We applied our method for profiling of Aβ peptides in CSF samples from nonAlzheimer patients and patients with Alzheimer’s disease. Aβ peptides in the CSF samples were captured and concentrated using a microfluidic system in which magnetic beads coated with anti-Aβ were self-organized into an affinity microcolumn under the a permanent magnetic field. Finally, we could detect two Aβ peptides (Aβ1-40 and Aβ1-42) in the CSF samples.