Title Page
ABSTRACT
Contents
ABBREVIATIONS 15
1. INTRODUCTION 16
2. EXPERIMENTAL 20
2.1. Reagents 20
2.2. Buffers 21
2.3. Instrumentation 21
2.4. Synthesis of the coating antigen (BSA-SDMA) 22
2.5. Optimization of assay conditions 23
2.6. Optimization of pre-incubation 23
2.7. Dose-response curve in buffer 24
2.8. Cross-reactivity study 25
2.9. Dose-response curve in serum 26
2.10. Recovery study 26
3. RESULTS AND DISCUSSION 28
3.1. Synthesis and optimization of the coating antigen (BSA-SDMA) 28
3.2. Optimization of assay conditions 29
3.2.1. Effects of assay buffer and wash buffer 29
3.2.2. Effect of pre-incubation 29
3.2.3. Optimization of binder 30
3.2.4. Optimization of signal generator(avidin-HRP) 30
3.3. Dose-response curve in buffer 31
3.4. Cross-reactivity study 32
3.5. Dose-response curve in serum 32
3.6. Recovery study 33
4. CONCLUSION 34
5. REFERENCES 37
요약 57
Table 1. Cross-reactivity (%) of cross reactants for SDMA by single-site immunometric assay. 54
Table 2. Recovery study for determination of SDMA spiked in serum. 55
Table 3. Analytical data for SDMA in this study and other analytical methods. 56
Figure 1. Chemical structures of SDMA and cross-reactants. 44
Figure 2. The affinity test of BSA-SDMA conjugates. The ratio of... 45
Figure 3. Optimization of concentration of BSA-SDMA conjugates. The... 46
Figure 4. Optimization of assay buffer and wash buffer. The buffer... 47
Figure 5. Optimization of pre-incubation time. Pre-incubation time was 0... 48
Figure 6. Optimization of concentration of anti-SDMA antibody-biotin.... 49
Figure 7. The affinity test of avidin-HRP concentration. The concentration... 50
Figure 8. Dose-response curve and calibration curve (inset) for SDMA in... 51
Figure 9. Cross-reactivity on optimized conditions of assay. The analytes... 52
Figure 10. Dose-response curve and calibration curve (inset) for SDMA in... 53
Scheme 1. A scheme for BSA-SDMA conjugates 42
Scheme 2. Schematic representation of single-site immunometric assay for SDMA detection. 43