Construction of a magnetic driven-quantum dots ratiometric fluorescence-immunosensor for rapid and accurate detection of C-reactive protein

Shenping Yu; Yangchao Shi; Yanbing Lv; Li Wang; Yuke Ma; Ning Li; Xiuyuan Zuo; Yifan Li; Jinjin Fan; Zifeng Zhang; Ruili Wu; Huaibin Shen; Lin Song Li

doi:10.26599/NR.2025.94907207

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Research Article | Open Access | Online First

Construction of a magnetic driven-quantum dots ratiometric fluorescence-immunosensor for rapid and accurate detection of C-reactive protein

Shenping Yu^{¹^,^§}, Yangchao Shi^{¹^,^§}, Yanbing Lv^¹

(), Li Wang^², Yuke Ma^², Ning Li^¹, Xiuyuan Zuo^³, Yifan Li^⁴

(), Jinjin Fan^¹, Zifeng Zhang^¹

(), Ruili Wu^¹, Huaibin Shen^¹, Lin Song Li^¹

()

1Key Lab for Special Functional Materials of Ministry of Education, and School of Nanoscience and Materials Engineering, Henan University, Kaifeng 475004, China

2Department of Clinical Laboratory, The First Affiliated Hospital of Henan University, Kaifeng 475004, China

3Liaoning Key Laboratory of Chemical Additive Synthesis and Separation, College of Chemistry and Environmental Engineering, Yingkou Institute of Technology, Yingkou 115014, China

4Department of Materials Science & Engineering, Iowa State University, Ames, Iowa 50011, USA

^§ Shenping Yu and Yangchao Shi contributed equally to this work.

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A novel immunosensor based on quantum dots (QDs) ratiometric fluorescence (QRF) and magnetic driven technology was developed for sensitive, rapid, accurate, and high-throughput detection of biomarkers. As a proof of concept, it has excellent performance in the detection of the inflammatory factor C-reactive protein (CRP) antigen.

Abstract

Efficient, sensitive, and convenient quantum dots (QDs) fluorescent immunosensors have a wide range of applications in the field of in vitro diagnostics (IVD). However, traditional single-signal readout methods are susceptible to interference, have limited sensitivity, and involve time-consuming detection processes. These have become key bottlenecks restricting their use in point-of-care (POC) detection. Herein, we developed a novel immunosensor based on QDs ratiometric fluorescence (QRF) and magnetic driven technology for sensitive, rapid, accurate detection of C-reactive protein (CRP). High-quality magnetic-fluorescent QDs nanobeads (QBs) were prepared by efficiently and orderly layered loading of magnetic nanoparticles (MNP) and red QDs using mesoporous silica nanoparticles (MSN) as a template. The magnetic driven-QRF-immunosensor was constructed based on green QDs probes-functionalized microplate. The results showed that the linear range of magnetic-fluorescence probes for the detection of CRP was 0.1–1000 ng/mL, with a sensitivity of up to 0.05 ng/mL. Compared to conventional QRF-immunosensor, the detection time was halved (within 30 min), and the sensitivity improved by approximately 5-fold. Recovery experiments and clinical sample analyses also demonstrated the good accuracy of this immunoassay. Therefore, the constructed novel magnetic driven-QRF-immunosensor shows great potential for clinical diagnosis and disease monitoring, and is expected to be applied in POC detection as a new generation of detection technology.

Keywords

magnetic-fluorescent quantum dots ratiometric fluorescence magnetic driven immunosensor rapid and accurate

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References

[1]

Wu, P.; Yan, X. P. Doped quantum dots for chemo/biosensing and bioimaging. Chem. Soc. Rev. 2013, 42, 5489–5521.

Material	Method	Range (ng/mL)	LOD	References
AuNP	Aptamer-based colorimetric assay	889–20,700	1.2 pg/mL	[41]
AuNPs@BP@PDA	Electrochemical detection	0–36	0.7 ng/mL	[42]
Graphene oxide	Electrochemical detection	0.01–100	3.3 pg/mL	[43]
QDs	LIFA	0.5–1000	0.3 ng/mL	[44]
QDs	QD-FLISA	1.56–400	0.46 ng/mL	[45]
N-GQDs/AuNPs	Fluorescent aptasensor	0.2–300	0.2 ng/mL	[46]
Nanoplasmonic based on the EoT	Ultrasensitive nanoplasmonic immunoturbidimetry assay	1–500	0.54 ng/mL	[47]
Hollow ruthenium nanoparticles	Colorimetric detection	0.12–7.8	33.2 pg/mL	[48]
AIEgen-doped nanobeads	LIFA	8–100	0.16 ng/mL	[49]
AuNP	Sandwich ELISA	0.1–200	8 pg/mL	[50]
Cu-MOF	Fluorescence assay	0.1–50	40 pg/mL	[51]
Magnetic-fluorescent QBs	Magnetic driven-QRF-immunosensor	0.1–1000	0.05 ng/mL	This work

Sample number	Experimental value (ng/mL)	Spiked value (ng/mL)	Recovery (%)	RSD (%)
1	5	4.83	96.5	4.9
2	50	51	102	4.1
3	500	502.1	100.4	6.2