학회 | 한국화학공학회 |
학술대회 | 2007년 봄 (04/19 ~ 04/20, 울산 롯데호텔) |
권호 | 13권 1호, p.400 |
발표분야 | 생물화공 |
제목 | Improving the productivity of scFv against c-Met by rearranging the order of variable domains |
초록 | c-Met, a high affinity receptor for hepatocyte growth factor/scatter factor, shown to be overexpressed in a variety of malignant cells, is a potential biomarker as well as a therapeutic target. Thus antibody specific for c-Met is expected to be efficiently employed in the clinical treatment or imaging of many cancer cells. scFvs against c-Met with two different domain orders, i.e. VL-linker-VH and VH-linker-VL, were expressed in the cytoplasm of E. coli trx/gor deleted mutant and their activities as well as their productivities were compared. The scFv with VH/VL orientation showed 5-10 times higher expression levels than those with VL/VH orientation. Coexpression of DsbC in the cytoplasm of E. coli increased the yield of functional fragment antibodies about more than 5 fold compared to the production of fragment antibodies without DsbC coexpression. The fragment antibodies fused with hexahistidine residues at the C-terminus of the recombinant proteins were purified by immobilized metal affinity chromatography (IMAC). Acknowledgement : This research was supported by a grant (04-RIS-024) from Regional Innovation System funded by the Ministry of Commerce, Industry and Energy, Republic of Korea |
저자 | 김유진, 허미애, 이선구 |
소속 | 부산대 |
키워드 | c-Met; scFv; diabody; DsbC |
원문파일 | 초록 보기 |