| 학회 |
한국화학공학회 |
| 학술대회 |
2003년 봄 (04/25 ~ 04/26, 순천대학교) |
| 권호 |
9권 1호, p.451 |
| 발표분야 |
생물화공 |
| 제목 |
Production of Single Chain Antibodies against Mycotoxin Deoxynivalenol in Recombinant Escherichia coli |
| 초록 |
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 λ-2, respectively. The variable heavy (VH) and variable light (VL) genes were combined by overlap-extension PCR using a flexible DNA linker 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 His6-tag was purified by IMAC. The kinetic analysis for interactions between ligand (Ag) and analyte (Ab) were carried out by the BIAcore system. 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. |
| 저자 |
최규호1, 서진호1, 손동화2, 조영진1, 민원기1
|
| 소속 |
1서울대, 2한국식품개발(연) |
| 키워드 |
deoxynivalenol; single chain antibody; chaperone; IMAC; BIAcore
|
| E-Mail |
, |
| VOD |
VOD 보기 |
