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基于金属依赖型 DNA zyme 辅助纳米机器的血清前列腺特异性抗原放大荧光检测。

Amplified fluorescence detection of serum prostate specific antigen based on metal-dependent DNAzyme assistant nanomachine.

机构信息

Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Shandong Normal University, Jinan 250014, PR China; Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, College of Chemistry and Chemical Engineering, Linyi University, Linyi 276005, PR China.

Shandong Provincial Key Laboratory of Detection Technology for Tumor Markers, College of Chemistry and Chemical Engineering, Linyi University, Linyi 276005, PR China.

出版信息

Anal Chim Acta. 2018 May 30;1008:96-102. doi: 10.1016/j.aca.2017.12.041. Epub 2018 Jan 10.

Abstract

An amplified fluorescence biosensing strategy for serum prostate specific antigen (PSA) was developed on the basis of DNAzyme. In presence of cofactor Zn, Zn -dependent DNAzyme could cleave the hairpin substrate probes which were dispersed in solution and generate remarkable fluorescent signal. Taking advantage of the magnetic beads as a carrier, one target protein could bring plentiful hairpin substrate probes on to the electrode through a sandwich structure (Ab/PSA/biotin-Ab). Moreover, during the cleavage process of as formed DNAzyme, DNAzyme did not be destroyed and could further react with other hairpin probes, then generated continuous fluorescent signal. Benefited by this amplified strategy, the limit of detection (LOD) was low to 0.05 ng mL, which was much lower than our previous reports. This method could be applied to detect different protein biomarkers in serum without corresponding aptamers by changing the corresponding antibodies and thus showed a remarkable prospect in clinical application.

摘要

基于 DNA zyme,建立了一种用于血清前列腺特异性抗原(PSA)的荧光生物传感放大策略。在辅助因子 Zn 的存在下,Zn 依赖性 DNAzyme 可以切割分散在溶液中的发夹底物探针,产生显著的荧光信号。利用磁珠作为载体,通过夹心结构(Ab/PSA/生物素-Ab),一个靶蛋白可以将大量的发夹底物探针带到电极上。此外,在形成的 DNAzyme 的切割过程中,DNAzyme 不会被破坏,并且可以进一步与其他发夹探针反应,从而产生连续的荧光信号。受益于这种放大策略,检测限(LOD)低至 0.05ng/mL,明显低于我们之前的报告。该方法可以通过改变相应的抗体,应用于检测血清中不同的蛋白质生物标志物,而无需相应的适体,因此在临床应用中具有广阔的前景。

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