武汉云克隆科技股份有限公司

Abstract：

This paper reports a sensitive method with electrochemical technique to detect various proteases, which can be used for the diagnosis of prostate cancer. For the proposed assay method, the working electrode is modified with the peptide probes for the target proteases. These probes contain the substrate sequence of target proteases, as well as the seed peptide sequence that can accelerate the misfolding of amyloid-beta. If there are proteases in the test solution, after protease cleavage of the substrate peptides, the distal seed peptide will be removed from the electrode surface. So, in the absence of proteases, the seed peptides can initiate and accelerate amyloid-beta misfolding on the electrode surface. Consequently, the formed aggregates strongly block the electron transfer of the in-solution electroactive species with the electrode, resulting in suppressed signal readout. Nevertheless, in the presence of proteases, enzyme cleavage may lead to greatly mitigated protein misfolding and evident signal enhancement. Since the contrast in signal readout between the two cases can be amplified by using the protein misfolding step, high sensitivity suitable for direct detection of proteases in serum can be achieved. These results may suggest the feasibility of our new method for the detection of a panel of proteases in offering detailed diagnosis of prostate cancer and a better treatment of the cancer.

摘要：

本文研究了一种采用电化学技术检测各种蛋白酶的方法，可用于前列腺癌的诊断。对于所提出的检测方法，工作电极用肽探针修饰为靶蛋白酶。这些探针包含靶蛋白酶的底物序列，以及加速淀粉样β折叠的种子肽序列。如果在测试溶液中存在蛋白酶，底物肽蛋白酶裂解后，电极表面上的远端种子肽将被去除。因此在没有蛋白酶的情况下，电极表面的种子肽可以诱导和加速淀粉样β错折叠。因此，形成的聚集体强烈地阻碍了溶液中电活性物种与电极的电子传递，从而抑制了信号读出。然而，在蛋白酶的存在下，酶的切割可能会大大减轻蛋白质的错折叠和明显的信号增强。因为蛋白质错折叠步骤可以放大两例之间的信号读出对比，可获得较高的灵敏度，适用于血清中蛋白酶的直接检测。研究结果显示出这种检测蛋白酶新方法的可行性，为前列腺癌详细诊断和癌症治疗提供依据。