研究目的
To construct an ultrasensitive and highly selective electrochemiluminescence immunosensor for the detection of prostate specific antigen (PSA) with a wide linear range and low detection limit, addressing the need for high sensitivity and wide linear range in PSA detection for clinical applications such as prostate cancer diagnosis and postoperative evaluation.
研究成果
The developed ECL immunosensor demonstrates high sensitivity with a wide linear range of 1 pg/mL to 150 ng/mL and a low detection limit of 0.6 pg/mL for PSA, attributed to the multiple amplification strategy using GOx and Pt NPs conjugated with PGA. It shows excellent selectivity and reproducibility, making it suitable for clinical applications in prostate cancer diagnosis and monitoring. The general sensing scheme can be adapted for detecting other biological molecules, indicating broad potential utility.
研究不足
The study may have limitations in terms of real-sample applicability, potential interference from complex biological matrices, and the need for optimization for other biomarkers. The stability and long-term performance of the immunosensor in clinical settings were not extensively discussed, and scalability for mass production might require further investigation.
1：Experimental Design and Method Selection:
The study employs a multiple amplification approach using glucose oxidase (GOx) and platinum nanoparticles (Pt NPs) conjugated with polyglutamic acid (PGA) to enhance the electrochemiluminescence signal for PSA detection. The immunosensor is designed with a glassy carbon electrode modified with Au NPs-luminol-LDH for antibody immobilization and signal generation.
2：Sample Selection and Data Sources:
PSA standard antigen solutions of different concentrations were used, along with control substances like BSA, human IgG, and cTnT to test selectivity. The samples were prepared and measured under controlled conditions.
3：List of Experimental Equipment and Materials:
Key materials include bovine serum albumin (BSA), N-(3-(dimethylamino)propyl)-N′-ethyl-carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), luminol, poly-glutamic acid (PGA), chloroplatinic acid (H2PtCl6·6H2O), prostate specific antigen, monoclonal anti-PSA antibody (Ab1), polyclonal anti-PSA antibody (Ab2), glucose oxidase (GOx), and others from suppliers like Sigma Aldrich, Macklin Inc., Sinopharm Chemical Reagent Co. Ltd., and Sangon Biotech Co. Ltd. Equipment includes an MPI-E ECL analyzer, Ag/AgCl reference electrode, platinum wire counter electrode, and centrifugation tools.
4：Experimental Procedures and Operational Workflow:
The procedure involves preparation of Ab2-GOx-PGA-Pt NPs probe using EDC/NHS conjugation method, immobilization of Ab1 on Au NPs-luminol-LDH modified electrode, blocking with BSA, incubation with PSA and the probe, and ECL measurement using the MPI-E analyzer in a buffer containing glucose. Steps include centrifugation, dispersion, and incubation at specific temperatures and times.
5：Data Analysis Methods:
ECL intensities were measured and correlated with PSA concentrations. Statistical analysis included calculation of linear range, detection limit (S/N=3), selectivity against controls, and reproducibility (RSD from multiple measurements).
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