title: A new method to assay protease based on amyloid misfolding: application to prostate cancer diagnosis using a panel of proteases biomarkers.
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.
authors.affiliation: 1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.affiliation: 1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Huang
authors.affiliation: 2. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.affiliation: 1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Yang
authors.affiliation: 2. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhu
authors.affiliation: 1. Department of Biochemistry and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China. ; 2. Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai, China.
authors.forename: Genxi
authors.initials: G
authors.lastname: Li
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry*; D016022:Case-Control Studies; D055088:Early Detection of Cancer / Q000379:methods*; D006801:Humans; D008297:Male; D017430:Prostate-Specific Antigen / Q000097:blood / Q000737:chemistry; D011471:Prostatic Neoplasms / Q000175:diagnosis*; D017510:Protein Folding; D059748:Proteolysis; D012697:Serine Endopeptidases / Q000097:blood / Q000737:chemistry*; D020840:Tissue Kallikreins / Q000097:blood / Q000737:chemistry*
chemical_list: D016229:Amyloid beta-Peptides; D012697:Serine Endopeptidases; C485595:matriptase; D020840:Tissue Kallikreins; D017430:Prostate-Specific Antigen
keywords: amyloid misfolding; biomarker; electrochemical detection.; prostate cancer; protease