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. 2023 Apr 19;23(8):4118.
doi: 10.3390/s23084118.

New Biosensor for Determination of Neuropilin-1 with Detection by Surface Plasmon Resonance Imaging

Anna Sankiewicz  1 , Beata Zelazowska-Rutkowska  2 , Ewelina Gorska  3   4 , Adam Hermanowicz  4 , Ewa Gorodkiewicz  1
Affiliations

New Biosensor for Determination of Neuropilin-1 with Detection by Surface Plasmon Resonance Imaging

Anna Sankiewicz et al. Sensors (Basel). .

Abstract

Neuropilin-1 is transmembrane protein with soluble isoforms. It plays a pivotal role in both physiological and pathological processes. NRP-1 is involved in the immune response, formation of neuronal circuits, angiogenesis, survival and migration of cells. The specific SPRI biosensor for the determination of neuropilin-1 was constructed using mouse monoclonal antibody that captures unbound NRP-1 form body fluids. The biosensor exhibits linearity of the analytical signal between 0.01 and 2.5 ng/mL, average precision value 4.7% and recovery between 97% and 104%. The detection limit is 0.011 ng/mL, and the limit of quantification is 0.038 ng/mL. The biosensor was validated by parallel determination of NRP-1 in serum and saliva samples using the ELISA test, with good agreement of the results.

Keywords: biosensor SPRi; neuropilin-1; saliva; serum.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Scheme of the sensor preparation procedure.
Figure 2
Figure 2
Dependence of SPRI signal (Arbitrary Units) on mouse NRP-1 antibody concentration. NRP-1 concentration: 5 ng/mL. pH = 7.4. Error bars were calculated for 12 independent measurements for each concentration at a 95%confidence level.
Figure 3
Figure 3
Dependence of the SPRI signal (Arbitrary Units) on NRP-1 concentration. (―): in the full range of tested concentrations, (―): calibration curve. NRP-1 antibody concentration: 200 ng/mL, pH = 7.4. Error bars were calculated for 12 independent measurements for each concentration at a 95% confidence level.
Figure 4
Figure 4
The experiment to test the selectivity of the biosensor.
Figure 5
Figure 5
QCM studies of the biosensors surface. Variation of frequency as a function of time.
Figure 6
Figure 6
Correlation between serum (A) and saliva (B) NRP-1 levels quantified with the SPRI-biosensor and the traditional ELISA assay. Each point represents a single patient. The plot of correlation (―), confidence interval ( - - - ).
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