| 초록 |
In this study we attempted to express the scFv antibodies that had been engineered to penetrate into the cells and give signal through split GFP complementation. Conventional cell-based gene expression methods often fail to produce sufficient amount of 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. scFv productivity was enhanced by adding a ubiquitin sequence in front of the ORF of scFv sequence. To increase the solubility, we used molecular chaperon GroEL/ES-enriched extract. To form correct disulfide bonds, we have changed redox conditions of the cell extract and reaction mixture using GSSG/GSH and DsbC. We expect that the established conditions would be applied as a general strategy for producing active scFvs. |
