아주대학교

목차

1. Introduction 1
1.1. Therapeutic antibodies 1
1.2. Antibody–drug conjugates (ADCs) 3
1.2.1. The structure and synthesis of ADCs 5
1.2.2. Mechanism of action of ADCs 7
1.2.3. Strategies to enhance antibody internalization 9
1.3. Bioorthogonal chemistry as a tool to control antibody multimerization 11
1.4. Aim of this study 12
2. Methods and Materials 14
2.1. Antibody expression and purification 14
2.2. Peptide-Directed Photo-Crosslinking (PEDIP) 15
2.3. Click conjugation of antibodies 16
2.4. Synthesis of ADC using lysine coupling 17
2.5. LC–MS analysis of antibody conjugates 18
2.6. Western blotting 19
2.7. Live-cell imaging 20
2.8. Confocal microscopy 21
2.9. xCELLigence real-time cell analysis (RTCA) cytotoxicity assay 22
3. Results and Discussion 23
3.1. Design of in situ multimerization platform 23
3.1.1. Synthesis of click conjugates 24
3.1.2. Demonstration of the in situ multimerization in the trastuzumab model 32
3.2. In situ multimerization promotes antibody internalization 36
3.3. In situ multimerization in heterotypic combinations 44
3.3.1. Synthesis of click conjugates 45
3.3.2. Enhanced Internalization via heterotypic multimerization 50
3.4. Application to antibody–drug conjugates (ADCs) 58
3.4.1. Synthesis and characterization of avelumab-based ADCs 59
3.4.2. Rapid and potent cytotoxicity of ADC via heterotypic in situ multimerization 62
4. Conclusion 67
5. References 69
6. Abstracts in Korean 71