Contents
摘要 I
Abstract II
誌謝 III
Contents IV
Figure contents VI
Table contents VIII
Abbreviations IX
Introduction 1
Antibodies 1
Phage 2
Phage display technology 3
Application of phage display 5
Biopanning 6
Antibody phage-displayed library 6
Thrombomodulin 7
Enterovirus 71 8
Study Design 10
Materials and Methods 11
Materials 11
Bacteria Strain 11
Helper phage 11
RNA isolation from Peripheral Blood lymphocytes 11
Reverse Transcription Reaction (RT) 12
Amplification of VH and VL repertoires 13
Overlapping PCR for producing the scFv repertoires 14
Electroporation 14
Rescue of phage library 15
Biopanning for recombinant thrombomodulin (rTM) 16
Biopanning for Enterovirus 71 (EV-71) 18
Colonies selected for binding screening 19
ELISA assay for binding screening 20
Purification of soluble rTM 3-9F scFv and EV 3-3H scFv 21
Western Blot 22
Results 24
Construction of scFv gene repertoires 24
Construction of phage scFv library 24
Biopanning of recombinant thrombomodulin (rTM) 26
Biopanning of Enterovirus 71 viral particles (EV-71) 27
Purification of soluble rTM 3-9F scFV 28
Purification of soluble EV3-3H scFv 28
Characterization of scFv-phage and scFvs for rTM and EV-71 29
Discussion 30
Conclusion 36
Reference 37
Figure 45
Table 70
Appendix 84
作者簡歷 88
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