| 학회 |
한국공업화학회 |
| 학술대회 |
2015년 가을 (11/04 ~ 11/06, 제주국제컨벤션센터(ICCJEJU)) |
| 권호 |
19권 2호 |
| 발표분야 |
생물공학_포스터 |
| 제목 |
Cell-free synthesis of disulfide bonds containing scFv with cell penetrating activity |
| 초록 |
Conventional cell-based gene expression methods often fail to produce sufficient amount of functional scFv. In contrast, cell-free protein synthesis system provides a high accessibility to the translational mechanism, thereby providing optimal environments to increase the productivity and solubility of scFv. In this study we attempted to express in a cell-free protein synthesis system scFv antibodies that had been engineered to penetrate into the cells. To increase the solubility, we used DsbA, DsbC, FkpA, SurA enriched extract. We also used a cleavable ubiquitin signal sequence in front of the target scFvs. To assist the formation of correct disulfide bonds, we have changed redox conditions of the cell extract and reaction mixture using GSSG/GSH redox buffer. Taken together, these modifications of the cell-free synthesis system allowed substantial increase in the productivity and solubility of the expressed scFvs. We expect that the established conditions would be applied as a general strategy for producing active scFvs. |
| 저자 |
김동명1, 유태현2, 김용성2, 최동기2, Devi Kasi1, Christy Catherine1, 민승의1, 이경호1
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| 소속 |
1충남대, 2아주대 |
| 키워드 |
cell-free protein synthesis. scFv; disulfide bonds formation
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| E-Mail |
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