| 학회 |
한국화학공학회 |
| 학술대회 |
2005년 가을 (10/21 ~ 10/22, 인하대학교) |
| 권호 |
11권 2호, p.2047 |
| 발표분야 |
생물화공 |
| 제목 |
Production and characterization of anti-TNF-α Single Chain Variable Fragment Antibody in Recombinant Escherichia coli |
| 초록 |
Tumor necrosis factor-α (TNF-α), a pleiotropic cytokine primarily produced by activated machrophage, became a useful target of therapy for several autoimmune diseases. Genes for the single chain variable fragment (scFv) of a monoclonal antibody (mAb) against TNF-α were cloned and expressed in Escherichia coli. Fusion of myeloma cells and spleen cells from the immunized mice resulted in hybridoma cells which were cultured to obtain the anti-TNF-α mAb. The complemetary DNA was constructed by the reverse transcription-polymerase chain reaction. 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 k. Overlap-extension PCR using a linker encoding polypeptide (Gly4Ser)3 led to combination of VH and VL genes. Anti-TNF-α scFv which was expressed insoluble in the recombinant E. coli was purified by using the poly Histidine tag and refolded in vitro. ELISA analysis determined the binding affinity of the refolded TNF-α scFv one third of the anti-TNF-α mAb. |
| 저자 |
봉기태, 민원기, 서진호
|
| 소속 |
서울대 |
| 키워드 |
TNF-a; scFv; recombinant E. coli; ELISA
|
| E-Mail |
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| 원문파일 |
초록 보기 |
