| 학회 |
한국화학공학회 |
| 학술대회 |
2004년 봄 (04/23 ~ 04/24, 공주대학교) |
| 권호 |
10권 1호, p.417 |
| 발표분야 |
생물화공 |
| 제목 |
Production of Single Chain Antibody against Mycotoxin Aflatoxin B1 in Recombinant Escherichia coli. |
| 초록 |
Aflatoxin B1 (AFB1) is a secondary metabolite produced mostly by Aspergillus flavus strain and toxic to human and several animal species as food contaminant. The single chain variable fragment (scFv) genes of a monoclonal antibody (mAb) against AFB1) were cloned and expressed in recombinant Escherichia coli. A fusion of myeloma cells and spleen cells from the immunized mice, hybridoma cells were cultured to obtain anti-AFB 1 mAb of 1.3 ng/ml IC50and 0.1 ng/ml detection limit. Complementary DNA was constructed by the reverse transcription-polymerase chain reaction and DNA sequence analysis identified that each variable region was composed of the heavy chain variable region (VH) as a type of IgG1 and the light chain variable region (VL) as a type of λ. Overlap-extension PCR using linker encoding polypeptide (Gly4Ser)3 led to combination of VH and VL genes and expression in recombinant E. coli. Co-expression of molecular chaperones enhanced soluble expression of anti-AFB1 scFv and the ubiquitin fusion system increased expression levels. In vitro refolding was performed and competitive direct enzyme linked immunosorbent assay determined the optimized condition. |
| 저자 |
민원기1, 조영진1, 최규호1, 권대혁1, 손동화2, 박경문3, 서진호1
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| 소속 |
1서울대, 2한국식품개발(연), 3유진사이언스 |
| 키워드 |
Aflatoxins B1; competitive direct enzyme linked immunosorbent assay; ubiquitin; molecular chaperones; hybridoma cell; single chain variable fragment
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| E-Mail |
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| 원문파일 |
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