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Production of single chain antibodies against mycotoxin deoxynivalenol in recombinant escherichia coli : 재조합 대장균을 이용한 곰팡이 독소 deoxynivalenol에 대한 특이적인 재조합 항체의 생산
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- Authors
- Advisor
- 서진호
- Issue Date
- 2003
- Publisher
- 서울대학교 대학원
- Keywords
- Deoxynivalenol ; Single chain antibody ; Chaperone ; Imac ; Biacore
- Description
- Thesis (master`s)--서울대학교 대학원 :농생명공학부,2003.
- Abstract
- The variable region genes of a monoclonal antibody specific for trichothecene
mycotoxin deoxynivalenol (DON) were cloned, characterized and expressed in
recombinant Escherichia coli. The sequence analysis revealed that the variable
regions belong to the mouse heavy chain subgroup I and light-chain subgroup ¥ë-2,
respectively. The variable heavy (VH) and variable light (VL) genes were
combined by overlap-extension PCR using a flexible DNA linker encoding
polypeptide (Gly4Ser)3 as single-chain variable fragment (scFv). In order to
express the funtional scFv antibody in recombinant E. coli, various expression
strategies were introduced. Coexpression of molecular chaperone (DnaK-DnaJ-GrpE)
with the anti-DON scFv antibody increased the soluble fraction of the scFv. The
scFv antibody containing the His 6-tag was purified by immobilized metal
affinity chromatography (IMAC). The kinetic analysis for interactions between
ligand (Ag) and analyte (Ab) were carried out by the BIAcore system using
surface plasmon resonance (SPR). The KD-value of the anti-DON scFv antibody was
found to be approximately 200-fold lower than that of the anti-DON monoclonal
antibody.
- Language
- English
- URI
- http://dcollection.snu.ac.kr:80/jsp/common/DcLoOrgPer.jsp?sItemId=000000058422
https://hdl.handle.net/10371/67620
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