Title Page
Contents
I. INTRODUCTION 8
II. MATERIAL AND METHODS 11
2.1. Materials and reagents 11
2.2. Optimized production of the CA-QCT inclusion complex 11
2.3. Inclusion product analysis by high-performance liquid chromatography (HPLC) and purification of CA-QCT 12
2.4. Identification of inclusion compounds 12
2.5. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS) 13
2.6. Analysis of solubility and stability of CA-QCT 13
2.7. Free-radical scavenging assay 14
2.8. Cell culture 14
2.9. Cell viability 15
2.10. Assay of nitric oxide production 15
2.11. Assay of interleukin-1β (IL-1β), interleukin-6 (IL-6), and cyclooxygenase-2 (COX-2) using a Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) 16
2.12. Statistical analysis 17
III. RESULTS 18
3.1. Enzyme reaction and purification of CA-QCT complex 18
3.2. Structural analysis of the CA-QCT complex 18
3.3. Physiochemical properties of CA-QCT in aqueous solution 22
3.4. Functional properties of CA-QCT complex 26
IV. DISCUSSION 32
V. REFERENCES 38
ABSTRACT 45
ABSTRACT KOREAN 46
Table 1. Theoretical molecular masses of CA-QCT complexes. 21
Table 2. Comparison of the water solubility between the QCT and QCT inclusion complexes 33
Figure 1. Preparation and purification of CA-QCT. 19
Figure 2. Analysis of the molecular weight of CA-QCT complexes using MALDI-TOF/MS. 20
Figure 3. Release of QCT from inclusion complexes after enzymatic hydrolysis. 23
Figure 4. Comparison of the stability of QCT and CA-QCT. 25
Figure 5. Comparison of the relative antioxidant effect of QCT and CA-QCT using a DPPH radical scavenging assay. 27
Figure 6. Effect of QCT and CA-QCT on RAW264.7 cell viability. 28
Figure 7. Inhibitory effect of QCT and CA-QCT on NO production in LPS-stimulated RAW264.7 cells. 29
Figure 8. Effects of QCT and CA-QCT on LPS-induced mRNA expression of IL-1β, IL-6, and COX-2. 31
Figure 9. Inclusion compound production at varying concentrations of CGTase. 37