Issue 24, 2023

Disposable capillary-fill device for the determination of proteases incorporating elimination of light-shielding from the magnetic beads with cleavage of the electrogenerated chemiluminescence label-tagged peptide probe

Abstract

A novel point-of-care testing (POCT) method for the determination of proteases was developed for the first time using a designed disposable capillary-fill device based on the cleavage of electrogenerated chemiluminescence (ECL)-label-tagged peptide probes and enabling elimination of the light-shielding from the magnetic beads (MBs). As a proof-of-principle, prostate-specific antigen (PSA) was taken as a model analyte, and streptavidin-coated magnetic beads bound with ruthenium-complex-tagged specific peptide (biotin-HSSKLQK) were utilized as MB ECL probes. The capillary-fill device was designed to be divided into a reaction zone and detection zone. In the reaction zone, the bio-cleavage reaction between the PSA analyte with the peptide on the surface of the MB ECL probes occurred, while in the detection zone, ECL emission was produced by a screen-printed carbon electrode, Ag/AgCl reference electrode and carbon counter electrode. When the analyte PSA was introduced into the suspension of MB ECL probes in the reaction zone of the device, biocleavage of the peptide occurred, and the cleaved Ru1 part was released from the surface of the MB ECL probes. The capillary-filled device was tilted 90°, and with the aid of gravity, the solution containing the released Ru1 part flowed to the surface of the working electrode in the detection region of the device, while the MB ECL probes were fixed in the reaction zone by an external magnet. PSA can be determined by the ECL emission from the released Ru1 part in the presence of the co-reactant tri-n-propylamine at the detection zone. Under the optimal conditions, the developed ECL method showed a low detection limit of 0.12 ng mL−1 for PSA. This work demonstrates that the developed ECL biosensing approach can eliminate the MB light-shielding effect and quantify proteases with high sensitivity and selectivity, which could be easily extended to POCT-based ECL biosensing for other proteases.

Graphical abstract: Disposable capillary-fill device for the determination of proteases incorporating elimination of light-shielding from the magnetic beads with cleavage of the electrogenerated chemiluminescence label-tagged peptide probe

Supplementary files

Article information

Article type
Paper
Submitted
16 Sep 2023
Accepted
27 Okt 2023
First published
29 Okt 2023

Analyst, 2023,148, 6253-6260

Disposable capillary-fill device for the determination of proteases incorporating elimination of light-shielding from the magnetic beads with cleavage of the electrogenerated chemiluminescence label-tagged peptide probe

X. Yang, J. Li, H. Qi, Q. Gao and C. Zhang, Analyst, 2023, 148, 6253 DOI: 10.1039/D3AN01591B

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements