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首页> 外文期刊>Analytica chimica acta >A new colorimetric platform for ultrasensitive detection of protein and cancer cells based on the assembly of nucleic acids and proteins
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A new colorimetric platform for ultrasensitive detection of protein and cancer cells based on the assembly of nucleic acids and proteins

机译：基于核酸和蛋白质组装的蛋白质和癌细胞超灵敏检测的新比色平台

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An amplified colorimetric method has been developed for the detection of protein and cancer cells based on the assembly of nucleic acids and proteins for the first time. In this process, the assembly of nucleic acids was triggered by a biotinylated DNA strand after a sandwich immunoreaction. The biotinylated DNA strand and sandwich immunocomplex were connected by streptavidin. Then, the assembly of biotinylated bovine serum albumin (Biotin-BSA) and streptavidin-horseradish peroxidase (SA-HRP) occurred at a node of the assembled products of nucleic acids through the biotin-streptavidin reaction. Under the catalysis of horseradish peroxidase, 3,3',5,5'-tetramethylbenzidine (TMB) was oxidized by H2O2 and the oxidized product was analyzed by its UV-vis absorbance signal and sensitive colorimetric detection. This colorimetric sensor could not only achieve the quantitative determination of protein by UV-vis absorbance but could also be applied for semiquantitative determination by digital visualization. Using alpha-fetoprotein (AFP) as the model target, this proposed colorimetric method showed a wide linear range from 5 pg/mL to 1 ng/mL with a detection limit of 1.95 pg/mL by the instrument, and even 5 pg/mL target protein could be distinguished simply by the naked eye. This approach was then expanded to detect cancer cells based on the recognition of folic acid receptors that were over-expressed on the cancer cells by folic acid-tethered DNA. More importantly, this strategy can be further used as a universal colorimetric method for the determination of viruses or other proteins by changing the corresponding antibodies. (C) 2015 Elsevier B.V. All rights reserved.
机译：首次开发了一种基于核酸和蛋白质组装的扩增比色法，用于检测蛋白质和癌细胞。在这个过程中，在三明治免疫反应后，核酸的组装由生物素化的DNA链触发。生物素化的DNA链和夹心免疫复合物通过链霉亲和素连接。然后，通过生物素-链霉亲和素反应，在核酸的组装产物的节点处发生了生物素化的牛血清白蛋白（Biotin-BSA）和链霉亲和素-辣根过氧化物酶（SA-HRP）的组装。在辣根过氧化物酶的催化下，H2O2将3,3'，5,5'-四甲基联苯胺（TMB）氧化，并通过紫外可见吸收信号和灵敏的比色检测法对氧化产物进行分析。该比色传感器不仅可以通过紫外可见吸收度对蛋白质进行定量测定，而且还可以用于通过数字可视化进行半定量测定。以甲胎蛋白（AFP）为模型目标，该拟议的比色法显示了从5 pg / mL到1 ng / mL的宽线性范围，仪器的检测极限为1.95 pg / mL，甚至5 pg / mL靶蛋白可以简单地用肉眼区分。然后基于对叶酸受体DNA过度表达于癌细胞上的叶酸受体的识别，将该方法扩展到检测癌细胞。更重要的是，该策略还可以用作通用比色法，通过更改相应的抗体来确定病毒或其他蛋白质。 （C）2015 Elsevier B.V.保留所有权利。

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