Multifunctional Encoded Self-Assembling Protein Nanofibrils as Platform for High-Throughput and Multiplexed Detection of Biomolecules

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• 作者：Ho-Man Chan ; Hung-Wing Li
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：December 15, 2011
• 年：2011
• 卷：83
• 期：24
• 页码：9370-9377
• 全文大小：919K
• 年卷期：v.83,no.24(December 15, 2011)
• ISSN：1520-6882

文摘

A one-dimensional nanofibrillar array formed by the co-assembly of native and biotin-functionalized beta-amyloid (A尾) peptide was developed for biomolecule sensing. With the presence of biotin moiety, a variety of biomolecular probes can be conjugated onto the nanofibrils, thus converting the protein assembly into a miniature biosensor. In this work, DNA probes were immobilized onto the fibril for the detection of cDNA sequences. The as-developed 鈥淒NA-nanoarray鈥?achieved a detection limit at subattomole level (183 fM in 10 渭L). This highly sensitive, yet simple, assay requires a trace amount of sample consumption (<10 渭L) and is pretreatment-free. In addition, we reported the preparation of alternate-segmented amyloid nanofibrils with multifunctionality. The fibrils hereby serve as an encoded template that can be visualized with various fluorescence labeling dyes for barcode recognition purpose, and, hence, multiplex detection of biomolecules was achieved. Regarding that each protein nanofibril represents a single detection platform, a large number of single fibrils simultaneously are monitored with the dual-color TIRFM in a high-throughput manner.