AJOU Central Library Repository: 다양한 물리화학적 환경에서 핵산 가수분해 능력을 가진 단일가닥 촉매 항체의 기능적 안정성 분석

Graduate School of Ajou University Department of Biomedical Sciences 3. Theses(Master)

다양한 물리화학적 환경에서 핵산 가수분해 능력을 가진 단일가닥 촉매 항체의 기능적 안정성 분석

Alternative Title: Functional stability of a nucleic acid-hydrolyzing single chain antibody in various biochemical and biophysical environments

Keyword: 3D8; nucleic-acid hydrolyzing activity; single chain variable fragment; functional stability; antiviral activity

Alternative Abstract: 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.

Appears in Collections:: Graduate School of Ajou University > Department of Biomedical Sciences > 3. Theses(Master)

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