Title Page
Contents
Abbreviation 7
Abstract 9
Introduction 11
Materials and methods 17
1.1. Materials 17
1.2. Expression of P9-CXCR4 17
1.3. Purification of P9D-CXCR4 18
1.4. ELISA analysis of CXCR4 antibodies 19
Results and discussion 20
1.5. P9D-CXCR4 Expression and purification 20
1.6. Specific interaction of 1A2 antibody to CXCR4 20
Conclusion 30
References 31
국문 초록 32
Figure 1. Structure and the function of CXCR4. (A) The 3D structure of CXCR4 (B) The path way of CXCR4 13
Figure 2. Disease free survival probability due to CXCR4. "Influence of CXCR4 overexpression on disease-free survival for triple-negative... 14
Figure 3. P9 sequence and the CXCR4 expressed using P9 protein. (A) The sequence of P9 protein... 15
Figure 4. APG-O and scheme of reconstituted P9D-CXCR4. Various amine groups were coupled with the carboxyl groups of poly-γ-glutamic acid... 16
Figure 5. Expression test of CXCR4. Cells cultured after induction for 3 h at 37℃ (I+37), 5 h at 25℃ (I+25), O/N at 18℃... 22
Figure 6. Ni-NTA purification of P9D-CXCR4. Cells cultured after induction for 3 h at 37℃ (I+37), 5 h at 25℃ (I+25), O/N at 18oC (I+18)... 23
Figure 7. Size exclusion chromatography of P9DCXCR4 using HiLoad Superdex200 16/600 PG. (A) The samples obtained from Ni-NTA was separated by using HiLoad Superdex200 16/600 PG.... 24
Figure 8. Concentration-dependent binding of 1A2 P9D-CXCR antibody to the immobilized chemokine receptor (CXCR4, GLP1R).... 25
Figure 9. Concentration-dependent binding of 1A2 P9D-CXCR antibody to the immobilized chemokine receptor (EP4, 5HT3A).... 26
Figure 10. Concentration-dependent binding of 1A2 P9D-CXCR antibody to the immobilized chemokine receptor (CB1, LPA1).... 27
Figure 11. Concentration-dependent binding of 1A2 P9D-CXCR antibody to the immobilized chemokine receptor (CCR7).... 28
Figure 12. The homology between CXCR4 and CCR7 amino acid sequence According to PDB the homology between the full length CXCR4 and CCR7 appeared to... 29