Development of an SPRi Immune Method for the Quantitative Detection of Osteopontin
Abstract
:Highlights
- SPRi biosensors for osteopontin determination have been developed.
- The bioreceptors are mouse or rabbit monoclonal anti-osteopontin antibodies attached to the gold surface of a chip via a cysteamine linker.
- The biosensors have sufficient sensitivity to determine osteopontin at levels characteristic of this protein in blood plasma without any signal enhancement.
Abstract
1. Introduction
2. Materials and Methods
2.1. Materials and Reagents
2.2. SPRi Measurement Methodology
2.3. Sensor Biofunctionalization
2.4. Sensor Regeneration
2.5. ELISA Measurements
3. Results and Discussion
3.1. Formation of Layers on the Biosensor Surface
3.2. Optimization of Experimental Parameters
3.2.1. Selecting the Bioreceptor Concentration
3.2.2. Optimization of Time Interaction
3.3. Estimating the Calibration Range
3.4. Precision, Accuracy, and Recovery of the SPRi Method for the Determination of Osteopontin
3.5. Interference Studies
3.6. Determination of Osteopontin Concentration in Biological Samples
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Biosensor with Mouse Antibody | Biosensor with Rabbit Antibody | |||||||
---|---|---|---|---|---|---|---|---|
C Added [ng mL−1] | C Found Mean [ng mL−1] | SD | Recovery [%] | CV [%] | C Found Mean [ng mL−1] | SD | Recovery [%] | CV [%] |
0.050 | 0.046 | 0.004 | 93 | 7.6 | 0.049 | 0.004 | 99 | 7.3 |
0.250 | 0.248 | 0.002 | 99 | 0.7 | 0.258 | 0.010 | 103 | 3.9 |
0.500 | 0.521 | 0.008 | 104 | 1.4 | 0.509 | 0.014 | 102 | 2.8 |
1.000 | 0.981 | 0.017 | 99 | 1.7 | 1.016 | 0.035 | 102 | 3.4 |
Bioreceptor | Nr Samples | OPN Concentration [ng mL−1] | |||
---|---|---|---|---|---|
Plasma Not Spiked with Osteopontin | Expected | Observed | Recovery [%] | ||
Mouse antibody | 1 | 12.0 | 16.0 | 15.9 | 99 |
2 | 12.6 | 16.6 | 17.1 | 103 | |
3 | 11.6 | 15.6 | 16.7 | 107 | |
4 | 10.3 | 14.3 | 13.5 | 94 | |
5 | 13.5 | 17.5 | 17.3 | 99 | |
6 | 18.7 | 22.7 | 21.1 | 93 | |
7 | 10.1 | 14.1 | 13.9 | 99 | |
8 | 15.3 | 19.3 | 20.2 | 105 | |
9 | 7.8 | 11.8 | 12.2 | 104 | |
Rabbit antibody | 1 | 12.8 | 16.8 | 15.6 | 93 |
2 | 13.1 | 17.4 | 17.9 | 105 | |
3 | 12.8 | 16.8 | 15.6 | 93 | |
4 | 11.5 | 15.5 | 14.4 | 93 | |
5 | 12.3 | 16.3 | 16.4 | 101 | |
6 | 17.9 | 21.9 | 20.7 | 95 | |
7 | 9.7 | 13.7 | 15.2 | 111 | |
8 | 15.4 | 19.4 | 17.9 | 92 | |
9 | 8.6 | 12.6 | 12.7 | 101 |
Potential Interferent | OPN Spike [ng mL−1] | Excess Over OPN | Found OPN (Mouse Antibody as Bioreceptor [ng mL−1] | Recovery [%] | Found OPN (Rabbit Antibody as Bioreceptor [ng mL−1] | Recovery [%] |
---|---|---|---|---|---|---|
Fibronectin | 0.50 | 1:10 | 0.514 ± 0.022 | 103 | 0.518 ± 0.011 | 102 |
0.50 | 1:100 | 0.501 ± 0.012 | 100 | 0.514 ± 0.018 | 103 | |
Collagen IV | 0.50 | 1:10 | 0.515 ± 0.023 | 103 | 0.519 ± 0.022 | 104 |
0.50 | 1:100 | 0.506 ± 0.034 | 101 | 0.506 ± 0.026 | 101 | |
Albumin | 0.50 | 1:100 | 0.531 ± 0.029 | 106 | 0.528 ± 0.017 | 106 |
0.50 | 1:1000 | 0.516 ± 0.038 | 103 | 0.518 ± 0.022 | 104 |
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Sankiewicz, A.; Żelazowska-Rutkowska, B.; Guszcz, T.; Gorodkiewicz, E. Development of an SPRi Immune Method for the Quantitative Detection of Osteopontin. Sensors 2025, 25, 3628. https://doi.org/10.3390/s25123628
Sankiewicz A, Żelazowska-Rutkowska B, Guszcz T, Gorodkiewicz E. Development of an SPRi Immune Method for the Quantitative Detection of Osteopontin. Sensors. 2025; 25(12):3628. https://doi.org/10.3390/s25123628
Chicago/Turabian StyleSankiewicz, Anna, Beata Żelazowska-Rutkowska, Tomasz Guszcz, and Ewa Gorodkiewicz. 2025. "Development of an SPRi Immune Method for the Quantitative Detection of Osteopontin" Sensors 25, no. 12: 3628. https://doi.org/10.3390/s25123628
APA StyleSankiewicz, A., Żelazowska-Rutkowska, B., Guszcz, T., & Gorodkiewicz, E. (2025). Development of an SPRi Immune Method for the Quantitative Detection of Osteopontin. Sensors, 25(12), 3628. https://doi.org/10.3390/s25123628