Earticle

초록

영어

Single chain Fv antibody fragment against c-Met (wt-VH-VL) poses loss of function because of poor structural stability due to reduction of disulfide bond while expressed in Escherichia coli cytoplasmic reducing condition. Previously we have reported its functional expression in a mutant E. coli strain containing cytoplasmic non-reducing condition, which lacks the genes for enzymes (gor, trx) breaking the disulfide bonds and favors the disulfide bond formation. But
by nature and in vitro evolution it has been found that there are structurally stable domains exists without disulfide bond. Therefore it was hypothesized and successfully demonstrated that it is possible to generate a structurally stable antibody fragments by grafting the complementary determining region (CDR) loops into a stable antibody framework structures. Here we successfully employed the same CDR-loop grafting methodology to generate a stable anti-c-Met chimeric scFv (ch-VH-VL) by grafting the CDR loops into a highly stable human antibody framework structure (H3k3). Comparative analysis of the predicted models of three dimensional structures of wt-VH-VL and ch-VH-VL showed local hydrophobic interaction pattern plays the key role in retaining internal network interactions of the fold of variable domains in the absence of
disulfide bond. Finally the observed key interaction pattern has been supported by the presence and absence of such pattern on available X-ray crystal structures of stable and unstable domains respectively.

저자

• Selvakumar Edwardraja [ Department of Chemical and Biochemical Engineering, Pusan National University ]
• Sun-Gu Lee [ Department of Chemical and Biochemical Engineering, Pusan National University ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576