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다양한 물리화학적 환경에서 핵산 가수분해 능력을 가진 단일가닥 촉매 항체의 기능적 안정성 분석
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 권명희 | - |
| dc.contributor.author | Lee joungmin | - |
| dc.date.accessioned | 2019-10-21T07:28:08Z | - |
| dc.date.available | 2019-10-21T07:28:08Z | - |
| dc.date.issued | 2016-08 | - |
| dc.identifier.other | 23026 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/18875 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명과학과,2016. 8 | - |
| dc.description.tableofcontents | I. INTRODUCTION 1 II. MATERIALS AND METHODS 5 A. Purification of 3D8 scFv from Escherichia coli 5 B. Enzyme-linked immunosorbent assay (ELISA) 5 C. Fluorescence resonance energy transfer (FRET)-based DNA cleavage assay 6 D. Dialysis 7 E. Agarose gel electrophoresis-based DNA cleavage assay 8 F. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) 8 G. Infection of H9N2 virus 8 H. Flow cytometry 8 III. RESULTS 11 A. Bacterial expression and purification of 3D8 scFv 11 B. Effect of temperature on functional stabilities of 3D8 scFv 13 C. Effect of pH on functional stabilities of 3D8 scFv 16 D. DNA-hydrolyzing activity of 3D8 scFv under various pH conditions for long-term storage 18 E. Functional stabilities of 3D8 scFv under reducing environment 21 F. Effect of lyophilization and freeze-thaw cycles on functional stability 23 G. Determination of unit for DNA-hydrolyzing activity of 3D8 scFv 25 H. Antiviral activity of 3D8 scFv depending on the unit 27 IV. DISCUSSION 30 V. CONCLUSION 33 REFERENCES 34 국문요약 39 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 다양한 물리화학적 환경에서 핵산 가수분해 능력을 가진 단일가닥 촉매 항체의 기능적 안정성 분석 | - |
| dc.title.alternative | Functional stability of a nucleic acid-hydrolyzing single chain antibody in various biochemical and biophysical environments | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Joungmin Lee | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2016. 8 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 758701 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000023026 | - |
| dc.subject.keyword | 3D8 | - |
| dc.subject.keyword | nucleic-acid hydrolyzing activity | - |
| dc.subject.keyword | single chain variable fragment | - |
| dc.subject.keyword | functional stability | - |
| dc.subject.keyword | antiviral activity | - |
| dc.description.alternativeAbstract | 3D8 single chain variable fragment (scFv) antibody has cell-penetrating activity, nucleic acid-binding activity, nucleic acid-hydrolyzing activity, and antiviral activity. As a first step toward development of antiviral reagent using 3D8 scFv, functional stability based on DNA-binding and -hydrolyzing activity of 3D8 scFv was analyzed under biochemical and biophysical conditions such as different temperature, pH, storage period, reducing environment, lyophilization, and repetitive freeze-thaw. The DNA-binding activity and -hydrolyzing activity were maintained for 6 h at 50°C. The optimal pH showing the DNA-binding activity and -hydrolyzing activity was exhibited for 2 h at pH 7-11. The DNA-binding and -hydrolyzing activity were fully retained in the presence of reducing agent, 2 mM dithiothreitol (DTT). Moreover, DNA-hydrolyzing activity of 3D8 scFv was maintained after lyophilization and 30 cycles of freeze-thaw. When the 3D8 scFv was stored in pH 3-12 for a month at 37°C, DNA-hydrolyzing activity was retained at pH 4-8. In addition, we defined the unit for DNA-hydrolyzing activity and investigated antiviral activity against influenza A virus according to the unit. Our study would provide the basic information in developing an antiviral reagent with 3D8 scFv. | - |
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- Graduate School of Ajou University > Department of Biomedical Sciences > 3. Theses(Master)
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