Aptamer-based bio-barcode assay for the detection of cytochrome-c released from apoptotic cells
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摘要The recently developed bio-barcode (BBC) assay using polymerase chain reaction (PCR) to generate signals has been shown to be an extraordinarily sensitive method to detect protein targets. The BBC assay involves a magnetic microparticle (with antibody to capture the target of interest) and gold nanoparticle (with recognition antibody and thiolated single-stranded barcode DNAs) to form a sandwich around the target. The concentration of target is determined by the amount of barcode DNA released from the nanoparticles. Here we describe a modification using aptamers to substitute the gold nanoparticles for the BBC assay. In this study, we isolated a 76-mer monoclonal aptamer against cytochrome-c (cyto-c) and this single-stranded DNA in defined 3D structure for cyto-c was used in the BBC assay for both recognition and readout reporting. After magnetic separation, the aptamer was amplified by PCR and this aptamer-based barcode (ABC) assay was sensitive enough to detect the cyto-c in culture medium released from the apoptotic cells after drug treatment at the picomolar level. When compared to the conventional cyto-c detection by Western blot analysis, our ABC assay is sensitive, and time for the detection and quantification with ready-made probes was only 3 h. (C) 2010 Elsevier Inc. All rights reserved.
著者Lau IPM, Ngan EKS, Loo JFC, Suen YK, Ho HP, Kong SK
期刊名稱Biochemical and Biophysical Research Communications
出版年份2010
月份5
日期14
卷號395
期次4
出版社Elsevier
頁次560 - 564
國際標準期刊號0006-291X
電子國際標準期刊號1090-2104
語言英式英語
關鍵詞Apoptosis; Aptamer; Bio-barcode assay; Cytochrome-c
Web of Science 學科類別Biochemistry & Molecular Biology; BIOCHEMISTRY & MOLECULAR BIOLOGY; Biophysics; BIOPHYSICS

上次更新時間 2020-16-10 於 23:59