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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제18권 제6호
- 발행연도
- 2008.1
- 수록면
- 1,185 - 1,190 (6page)
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Single-chain Fv (scFv) antibody against c-Met is expected
to be employed in clinical treatment or imaging of cancer
cells owing to the important biological roles of c-Met in
the proliferation of malignancies. Here, we show that the
productivity of scFv against c-Met in Escherichia coli is
significantly influenced by the orientation of its variable
domains. We generated anti-c-Met scFv antibodies with
two different domain orders (i.e., VL-linker-VH and VH-
linker-VL), expressed them in the cytoplasm of E. coli trx/
gor deleted mutant, and compared their specific activities
as well as their productivities. Productivity of total and
functional anti-c-Met scFv with VH/VL orientation was
more than five times higher than that with VL/VH format.
Coexpression of DsbC enhanced the yield of soluble
amounts of anti-c-Met scFv protein for both constructs. The
purified scFv antibodies of the two different formats
exhibited almost the same antigen-binding activities. We
also compared the productivities and specific activities of
anti-c-Met diabodies with VH/VL or VL/VH formats and
obtained similar results to the case of scFv antibodies.
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참고문헌 (17)
참고문헌 신청
Arndt, M. A. E. / 2004 / Antigen binding and stability properties of non-covalently linked anti- CD22 single-chain Fv dimers / FEBS Lett 578 : 257 ~ 261
Baneyx, F. / 1999 / Recombinant protein expression in Escherichia coli. Curr. Opin / Biotechnol 10 : 411 ~ 421
Bessette, P. H. / 1999 / Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm. Proc. Natl. Acad / Sci 96 : 13703 ~ 13708
Fernandez, L. A. / 2004 / Prokaryotic expression of antibodies and affibodies. Curr. Opin / Biotechnol 15 : 364 ~ 373
Hamilton, S. / 2001 / Improved production by domain inversion of single-chain Fv antibody fragment against high molecular weight proteoglycan for the radioimmunotargeting of
Baneyx, F. / 1999 / Recombinant protein expression in Escherichia coli. Curr. Opin / Biotechnol 10 : 411 ~ 421
Bessette, P. H. / 1999 / Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm. Proc. Natl. Acad / Sci 96 : 13703 ~ 13708
Fernandez, L. A. / 2004 / Prokaryotic expression of antibodies and affibodies. Curr. Opin / Biotechnol 15 : 364 ~ 373
Hamilton, S. / 2001 / Improved production by domain inversion of single-chain Fv antibody fragment against high molecular weight proteoglycan for the radioimmunotargeting of
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