Title Page
Contents
Ⅰ. Introduction 10
Ⅱ. Materials and Methods 13
1. Cell lines and culture media. 13
2. Expression and purification of ScFv-Fc antibodies. 13
3. Lentivirus transfection. 13
4. RNA interference Knock down. 13
5. Cell viability assay. 14
6. Colony-formation assay. 14
7. Wound-healing assay. 14
8. Immunoblotting. 15
9. Flow cytometry. 15
10. Immunoprecipitation and Liquid chromatography–mass spectrometry. 15
Ⅲ. Results 16
1. Screening ScFv antibody against G0/G1 cell cycle arrest. 16
2. HT-9 Ab inhibits proliferation and migration in HT29 cells. 18
3. HT-9 Ab inhibits proliferation and migration in solid cancer cells. 20
4. HT-9 Ab arrest G0/G1 phase in solid cancer cells. 24
5. HT-9 Ab induced apoptosis in solid cancer cells. 27
6. Identification of the target protein. 30
Ⅳ. Discussion 33
Ⅴ. Reference 35
Ⅵ. Abstract 40
Ⅶ. Abstract in Korean 41
Figure 1. Sub-cloning of ScFv library genes into lentiviral vector. 17
Figure 2. HT-9 Ab suppress the growth of HT29 cells. 19
Figure 3. HT-9 Ab has anti-proliferative effects on solid cancer cells. 21
Figure 4. HT-9 Ab inhibited the colony formation and migration in solid cancer cells. 23
Figure 5. HT-9 Ab induced G0/G1 phase arrest in solid cancer cells. 26
Figure 6. HT-9 Ab induced apoptosis in solid cancer cells. 29
Figure 7. Identification of a target protein recognized by HT-9 antibody. 31
Figure 8. Knockdown of RACK1 suppresses the expression of RACK1 in solid cancer cells. 32