Development of a repebody for targeting and intracellular delivery of proteins = 세포 내 단백질을 표적하며 전달할 수 있는 리피바디의 개발

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Probing protein-protein interactions in living cells is crucial for understanding the protein functions and drug development. Small-sized protein binders are considered effective and useful for such analysis. Here we describe the development and use of a repebody, which is a protein binder composed of LRR (Leucine-rich repeat) modules, for tracking protein-protein interaction and localization in real-time. We developed an anti-mRFP repebody through a phage display method. Moreover, we designed an efficient and an EGFR-specific delivery platform using repebody to transport protein into the cells. In Chapter 1, a repebody with high affinity for a red fluorescent protein was selected through a phage display, fused with a green fluorescent protein, and applied for tracing a red fluorescent protein-fused target protein in mammalian cells. We have also demonstrated that a repebody can be effectively used for the analysis of protein-protein interaction in living cells by live-cell imaging. Chapter 2 describes the development of EGFR-specific repebody based delivery platform for receptor-specific intracellular delivery of cargos. We have shown that intracellular delivery of green ?uorescent protein (GFP) and gelonin toxin into the cytoplasm of mammalian cells. In consistent with an expression level of EGFR, repebody delivery system showed green fluorescence and cytotoxicity effects in mammalian cells in a dose dependent manner. In this study, we successfully developed a repebody for targeting intracellular protein and repebody based delivery platform in living cells. Our results suggest that the developed repebodies can be widely used for the analysis of protein-protein interaction and an understanding of complex biological processes in living cells. Furthermore, the present approach of repebody immunotoxin can be a new therapeutic agent for the treatment of cancer.
Advisors
Kim, Hak Sungresearcher김학성researcher
Description
한국과학기술원 :나노과학기술대학원,
Publisher
한국과학기술원
Issue Date
2017
Identifier
325007
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 나노과학기술대학원, 2017.8,[v, 61 p. :]

Keywords

Repebody▼aProtein-protein interactions▼aIntracellular antibody▼aProtein delivery▼aBacterial toxin; 리피바디▼a단백질-단백질 상호작용▼a세포 내 인공항체▼a단백질 전달▼a세균 독소

URI
http://hdl.handle.net/10203/241745
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=718805&flag=dissertation
Appears in Collection
NT-Theses_Ph.D.(박사논문)
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