Electrogenerated Chemiluminescence Peptide-Based Biosensor for the Determination of Prostate-Specific Antigen Based on Target-Induced Cleavage of Peptide

Abstract

A novel electrogenerated chemiluminescence peptide-based biosensor (ECL-PB) for the determination of prostate-specific antigen (PSA) was developed on the basis of target-induced cleavage of a specific peptide within Nafion film incorporated with gold nanoparticles (AuNPs) and ECL emitting species. A specific peptide (CHSSKLQK) was used as a molecular recognition element; tris(2,2'-ripyridine) dichlororuthenium(II) (Ru(bpy)3(2+)) was used as ECL emitting species, and ferrocene carboxylic acid (Fc) was employed as ECL quencher. The ECL-PB biosensor was fabricated by casting the mixture of Nafion and AuNPs onto the surface of glassy carbon electrode to form AuNPs/Nafion film, and then, Ru(bpy)3(2+) was electrostatically adsorbed into the AuNPs/Nafion film; finally, the peptide-tagged with ferrocene carboxylic acid (Fc-peptide) was self-assembled onto the surface of the AuNPs. When PSA was present, it specifically cleaved the Fc-peptide, leading the quencher to leave the electrode and resulting in the increase of the ECL intensity obtained from the resulted electrode in 0.1 M phosphate buffer saline (pH 7.4) containing tri-n-propylamine. The results showed that the increased ECL intensity was directly linear to the logarithm of the concentration of PSA in the range from 5.0 × 10(-12) to 5.0 × 10(-9) g/mL. An extremely low detection limit of 8 × 10(-13) g/mL was achieved because of the signal amplification through AuNPs and the ECL background suppression through Fc as ECL quencher. This work demonstrates that the combination of the direct transduction of peptide cleavage events with the highly sensitive ECL method is a promising strategy for the design of enzymatic cleavage-based ECL biosensors with high sensitivity and selectivity.

Electrogenerated Chemiluminescence Bioanalytic System Based on Biocleavage of Probes and Homogeneous Detection

“…8 Bioanalytic sensing systems are frequently used to assay the proteases, based on biocleavage of the probes, which consist of a specific substrate peptide and label, such as radioactive, 9 fluorescent, 10−12 chemiluminescence, 13 electrochemical, 14 and ECL reagents. 15 There are two approaches that include biosensors and biosensing systems, for quantifying proteases. In the first approach, the biosensor was fabricated by directly immobilizing the probes (electroactive, 16,17 ECL reagent 18 labeled peptide substrates) on the surface of electrodes.…”

Peptide-based biosensor for the prostate-specific antigen using magnetic particle-bound invertase and a personal glucose meter for readout

“…[11] and chemically synthesized and obtained from Shanghai Apeptide Co., Ltd. All chemicals were of analytical reagent grade and were used as received. Doubly distilled water was used throughout the experiments.…”

“…Denmeade identified a specific peptide with the amino acid sequence HSSKLQ that can be used as a substrate to determine PSA with high specificity and stability [10]. Recently, several peptide-based biosensors for the detection of PSA have been developed [2,[11][12][13]. In this contribution, we constructed a gold nanoparticle based analytical protocol for the detection of PSA.…”

Detection of influenza virus by a biosensor based on the method combining electrochemiluminescence on binary SAMs modified Au electrode with an immunoliposome encapsulating Ru (II) complex

“…An immunosensor based on ECL has been constructed by combining the Au-NPs/MWCNTimmobilized GC electrode and dendrimer having Ru complex-modified DNA, and shows a detection limit of 4.2 fg/mL [14]. An ECL peptide-based biosensor for determination of PSA has successfully used the induced cleavage of peptide to gain high sensitivity; the lower detection limit is 8 × 10 −13 g/mL [15]. Liposomes are a promising tool for enhancing the sensitivity or for affording the site of various reactions.…”