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Screening of cell-penetrating antibodies using a cell-free protein synthesis system
( Christy ),( Devi Kasi ),이경호,박유진,박성욱,김용성,김동명 한국공업화학회 2016 한국공업화학회 연구논문 초록집 Vol.2016 No.0
With its unique open reaction format, cell-free protein synthesis provides a versatile platform for production and screening of heterologus recombinant proteins. In this study, we used the cell-free protein synthesis system for expression screening of cell-penetrating scFv antibodies from a designed mutant library. By streamlining cell-free expression of the library with a cell-based split GFP complementation assay, we could examine the cell-penetration efficiency and stability of individual mutant scFvs in a fast and parallel manner. Cell-free protein synthesis system enables direct access and control over translational conditions, which was proven to be advantageous in high-throughput screening of antibodies.
이경호,( Christy Catherine ),( Devi Kasi ),이승원,신현일,김유정,주정원,김동명 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.0
Screening of suitable antigen is an important step in the development of diagnostic and therapeutic tools in the medical field. This study describes high-throughput expression screening of parasite based on the techniques of cell-free protein synthesis. The libraries of Plasmodium vivax and Clonorchis sinensis genes were PCR-amplified and immobilized on streptavidin agarose beads. Through dual functionalization of the agarose beads to capture 6xhistidine-tagged proteins as well as the biotinylated PCR products, when expressed in a reaction mixture for cell-free synthesis,template DNA and the expressed protein were in situ immobilized on the same beads. This approach of in situ expression and isolation enables streamlined recovery and analysis of cell-free synthesized proteins and also allows facile identification of the genes coding antigenic proteins through direct PCR of the microbead-bound DNA.