Title Page
Contents
국문초록 10
ABSTRACT 12
I. Introduction 14
1. Theoretical background 18
A. Exosomes 18
B. Epithelial cell adhesion molecule 20
C. Polydiacetylene 21
2. Research purpose 22
II. Material and methods 23
1. Materials 23
2. Isolation of cancer cell-derived exosomes from cell culture medium 24
3. Preparation of an anti-EpCAM aptamer-polydiacetylene 25
A. Synthesis of NHS-modified diacetylene monomers 25
B. Synthesis of an anti-EpCAM aptamer-conjugated diacetylene monomers 25
C. Fabrication of an anti-EpCAM aptamer-polydiacetylene 26
4. Colorimetric detection of cancer cell-derived exosomes 27
5. Detection of cancer cell-derived exosomes in clinical human serum 28
6. Characteristic analysis of an anti-Epcam aptamer-polydiacetylene 29
III. Results 30
1. Characterization of cancer cell-derived exosomes 30
2. Colorimetric detection of cancer cell-derived exosomes 33
3. Selectivity detection toward cancer cell-derived exosomes 37
4. Morphological analysis of an anti-EpCAM aptamer-polydiacetylene 39
5. Detection of cancer cell-derived exosomes in human serum 41
IV. Discussion 43
V. Conclusion 45
VI. References 46
Table 1. Measurement of cancer cell-derived exosomes in human serum via the spiked and recovery method 42
Figure 1. (a) Preparation of anti-EpCAM Apt-PDA. (b) Colorimetric detection of cancer cell-derived exosomes by specific interactions... 17
Figure 2. Composition and various surface markers of exosomes. 19
Figure 3. EpCAM expression level in epithelial cancers. 20
Figure 4. Synthesis process and the color transition from blue-to-red by external stimuli of PDA. 21
Figure 5. Isolation of exosomes obtained from cell culture medium. 24
Figure 6. Characterization of cancer cell-derived exosomes isolated from HT-29 cells. (a) TEM image (Scale bar: 200 nm), (b) NTA analysis,... 31
Figure 7. FACS analysis of EpCAM expression in normal cell-derived exosomes isolated from WI-38 cells. 32
Figure 8. Optimization of anti-EpCAM aptamer-linked diacetylene monomers (TCDA-Apt) composition of anti-EpCAM Apt-PDA by cancer... 33
Figure 9. Optimization of reaction conditions of the anti-EpCAM Apt-PDA with a 10% ratio of TCDA-Apt. UV-vis absorbance spectra of... 34
Figure 10. Photochromic response of anti-EpCAM Apt-PDA with cancer cell-derived exosomes. (a) Photographs, (b) UV-vis absorbance spectra,... 36
Figure 11. Selectivity of the anti-EpCAM Apt-PDA. (a) Photographs and (b) CR(%) of the anti-EpCAM Apt-PDA after the addition of target... 38
Figure 12. Morphological characteristics of the anti-EpCAM Apt-PDA. SEM images of the anti-EpCAM Apt-PDA (a) before and (b) after the... 40