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Development and Characterization of Monoclonal Antibody Specific to Oncogenic Variant AIMP2-DX2

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dc.contributor.advisor김성훈-
dc.contributor.author성준식-
dc.date.accessioned2017-07-19T11:09:19Z-
dc.date.available2017-07-19T11:09:19Z-
dc.date.issued2016-08-
dc.identifier.other000000136125-
dc.identifier.urihttps://hdl.handle.net/10371/133403-
dc.description학위논문 (석사)-- 서울대학교 융합과학기술대학원 : 바이오제약학과, 2016. 8. 김성훈.-
dc.description.abstractAIMP2-DX2 is known as an exon2-deleted splicing variant of AIMP2 (amonoacyl-tRNA synthetase-interacting multifunctional protein 2) showing high expression in lung cancer cells and patients tissues. AIMP2-DX2 inhibits a pro-apoptotic role of AIMP2, which is known as an tumor suppressor, through competing for the association with common binding partners such as p53, FBP (fuse-binding protein) and TRAF2 (TNF receptor-associated factor 2) under various stimuli and stresses. According to previous studies, the cells with higher level of AIMP2-DX2 tend to show increased resistance to cell death. Also, suppression of AIMP2-DX2 in xenograft mouse model significantly retarded tumor growth.
Due to lack of monoclonal antibody specific to AIMP2-DX2, there has been a difficulty in developing diagnostic tools to classify patients with relatively higher level of AIMP2-DX2 for targeted cancer therapy. In this study, phage display technique was used to create monoclonal antibody that specifically binds to AIMP2-DX2. The newly generated monoclonal antibody was also validated and characterized by immunoblot, enzyme-linked immunosorbent assay, immunoprecipitation, surface plasmon resonance assay and immunofluorescence microscopy.
The results showed that the selected monoclonal antibody clone, H5, could specifically recognize AIMP2-DX2 but not AIMP2 and it showed a prominent binding interaction with the epitope peptide as well as purified AIMP2-DX2 protein. Based on these evidences, I concluded that the newly generated monoclonal antibody hold a great potential in both research and clinical fields and it could be employed in cancer diagnosis at in various ways.
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dc.description.tableofcontentsINTRODUCTION 7

MATERIALS AND METHODS 10
Rabbit Immunization, mRNA Extraction, and cDNA Synthesis 10
Generation of Rabbit Fab Fragment 11
Construction of Fab Antibody Library 11
Panning of Phage Displayed Antibody Library 12
Screening for Phage Library Clones 12
Sequencing of the Selected Clone 13
Cell Culture 14
Immunoblotting 14
Immunoprecipitation (IP) 15
Small Interfering RNA (siRNA) 15
Surface Plasmon Resonance (SPR) 16
Immunofluorescence (IF) 16

RESULTS 18
Rabbit immunization with AIMP2-DX2 Peptide Antigen 18
Construction and Selection of Monoclonal Antibody Library from Immunized Rabbit 19
Identification of Possible AIMP2-DX2 Isoforms 19
Conversion of H5 Fab into IgG1 Form 21
Characterization of H5 Monoclonal Antibody by Measuring the Affinity and Specificity against AIMP2-DX2 21

DISCUSSION 33

REFERENCES 36

요약 (국문초록) 39
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dc.formatapplication/pdf-
dc.format.extent1347385 bytes-
dc.format.mediumapplication/pdf-
dc.language.isoen-
dc.publisher서울대학교 융합과학기술대학원-
dc.subjectAIMP2-DX2-
dc.subjectMonoclonal Antibody-
dc.subjectLung cancer-
dc.subjectDiagnosis-
dc.subjectPhage Display-
dc.subject.ddc610-
dc.titleDevelopment and Characterization of Monoclonal Antibody Specific to Oncogenic Variant AIMP2-DX2-
dc.typeThesis-
dc.description.degreeMaster-
dc.citation.pages40-
dc.contributor.affiliation융합과학기술대학원 분자의학 및 바이오제약학과-
dc.date.awarded2016-08-
Appears in Collections:
Graduate School of Convergence Science and Technology (융합과학기술대학원)Dept. of Molecular and Biopharmaceutical Sciences (분자의학 및 바이오제약학과)Theses (Master's Degree_분자의학 및 바이오제약학과)
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