Earticle

초록

영어

Single chain Fv antibody (scFv) against c-Met is expected to be employed in clinical treatment or imaging of cancer cells due to the important biological roles of c-Met in the proliferation of malignancies. Hence it would be of importance to obtain a large amount of the recombinant anti-c-Met scFv for its practical utilization. Here, we show that the productivity of scFv against c-Met 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 amount of anti-c-Met scFv protein for both constructs. The scFv antibodies of the two different formats were purified through the interaction between Ni2+ and histidine tag on the c-terminus of recombinant protein, exhibiting almost 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 with the case of scFv antibodies.

저자

• Yu-Jin Kim [ Department of Chemical and Biochemical Engineering, Pusan National University ]
• Mi-Ae Heo [ Department of Chemical and Biochemical Engineering, Pusan National University ]
• Rameshkumar Neelamegam [ Department of Chemical and Biochemical Engineering, Pusan National University ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576