Title Page
Contents
LIST OF ABBREVIATIONS 11
ABSTRACT 13
Ⅰ. INTRODUCTION 16
1. Biosensor Market Trend 16
2. Biosensor Technology Trend 20
3. Summary of SARS-CoV-2 23
4. Wound and Wound Healing Stage Detection 28
5. Research Objectives 39
Ⅱ. MATERIALS AND METHODS 42
1. Electrochemical Biosensor 42
1.1. N Protein of SARS-CoV-2 Detection 42
2. Colorimetric Biosensor 44
2.1. MMP-9 Detection 44
2.2. Nitrite Ion Detection 47
2.3. pH Sensing 49
Ⅲ. RESULTS AND DISCUSSION 51
1. Electrochemical Biosensor 51
1.1. N Protein of SARS-CoV-2 Detection 51
2. Colorimetric Biosensor 73
2.1. MMP-9 Detection 73
2.2. Nitrite Ion Detection 83
2.3. pH Sensing 91
2.4. Multiple Sensors for Wound/Healing Detection 95
Ⅳ. CONCLUSION 97
Ⅴ. REFERENCES 101
ABSTRACT IN KOREAN 120
Table 1. Summary of the electrochemical SARS-CoV-2 detection methods 26
Table 2. Electrochemical and colorimetric biosensors advantages and disadvantages 38
Figure 1. In vitro diagnostics market prospects and market share 19
Figure 2. Technology classification for electrochemical and stimuli-responsive polymers 22
Figure 3. Schematic diagram of (A) The 3D model of the SARS-CoV-2 (B) The structure of the SARS-CoV-2 25
Figure 4. Illustration of four phases in the wound healing process 30
Figure 5. A summary of development for electrochemical and colorimetric biosensors 41
Figure 6. Operation of the impedance analyzer system. The system consists of an IWE sensor, impedance reader and software. During... 52
Figure 7. SEM image of the IWE. (a) The overall SEM image of an electrode. The electrode consists of a sensing area and contact pads... 53
Figure 8. Comparable to El-ISA and ELISA. El-ISA measures impedance in real time, whereas ELISA determines the optical density... 55
Figure 9. The El-ISA stabilization process. Admittance obviously changed in the first stabilization step but changed minimally (138~139... 57
Figure 10. Measurement of anti-N protein detection Ab concentration. (a) Admittance monitored with 0 to 80 ng/mL (b) Admittance... 60
Figure 11. Optimization of the blocking buffer for the El-ISA. Admittance of residual detection antibody was monitored for 1 h... 64
Figure 12. Principle of the El-ISA. A higher concentration of N protein results in less residual detection antibody and consequently... 64
Figure 13. Quantification of the residual detection antibody. (a) Admittance of the residual detection antibody monitored in solution... 65
Figure 14. Specificity of N protein between N and S proteins for El-ISA. (a) Changes in admittance of S (1 ng/mL) and N (0, 1, 5,... 67
Figure 15. Specificity of the N protein in the N and S protein mixture for El-ISA. Normalized admittance using a mixed solution of N and... 68
Figure 16. Specificity of N protein in a 20% human serum-containing solution for El-ISA. (a), (b). Concentrations of spiked N protein: 0,... 69
Figure 17. Specificity of N protein between N and S proteins for FIA. (a) N and S proteins determined by the FIA. (b) Fluorescence... 71
Figure 18. Specificity of N protein in a 20% human serum-containing solution for FIA. (a), (b) FIA in 20% human serum spiked with... 72
Figure 19. Level of MMP-9 protein in clinical samples. (A) saline diluted exudates and (B) undiluted exudates obtained from human... 75
Figure 20. Preparation of FRET-peptide-bound hydrogel patterns 77
Figure 21. The scheme of the MMP-9 sensing principle. The MMP-9 generates an increase in fluorescence. 78
Figure 22. Intensity of hydrogel sensors exposed to MMP-9 for 20 min. (a) Images acquired after a 20 min incubation with MMP-9... 80
Figure 23. Intensity of hydrogel sensors exposed to MMP-9 for 50 min. Plot of MMP-9 concentration vs. fluorescence intensity with 50... 81
Figure 24. Compare to ELISA data (Figure 19) and hydrogel sensor data. (a) Fluorescence images in clinical sample. (b) Plot of time vs.... 82
Figure 25. Nitrite sensor fabrication process 85
Figure 26. Color change of a glass fiber-type sensor in a urine sample 86
Figure 27. RGB extract data from 10 μM to 10 mM (a) and a 10 mM calculation example (b). 88
Figure 28. Color change of a hydrogel sheet-type sensor in urine and DI samples. 89
Figure 29. RGB data extracted from a hydrogel-based sensor in urine and DI samples. 90
Figure 30. Preparation of a pH sensor using a universal indicator. 92
Figure 31. pH sensor color change in 1 to 12 levels and RGB extract data. 94
Figure 32. Multiple sensors for wound and healing detection 96