RRC ID |
52142
|
著者 |
Nguyen TD, Takasuka H, Kaku Y, Inoue S, Nagamune T, Kawahara M.
|
タイトル |
Engineering a growth sensor to select intracellular antibodies in the cytosol of mammalian cells.
|
ジャーナル |
J Biosci Bioeng
|
Abstract |
Intracellular antibodies (intrabodies) are expected to function as therapeutics as well as tools for elucidating in vivo function of proteins. In this study, we propose a novel intrabody selection method in the cytosol of mammalian cells by utilizing a growth signal, induced by the interaction of the target antigen and an scFv-c-kit growth sensor. Here, we challenge this method to select specific intrabodies against rabies virus nucleoprotein (RV-N) for the first time. As a result, we successfully select antigen-specific intrabodies from a naïve synthetic library using phage panning followed by our growth sensor-based intracellular selection method, demonstrating the feasibility of the method. Additionally, we succeed in improving the response of the growth sensor by re-engineering the linker region of its construction. Collectively, the described selection method utilizing a growth sensor may become a highly efficient platform for selection of functional intrabodies in the future.
|
巻・号 |
124(1)
|
ページ |
125-132
|
公開日 |
2017-7-1
|
DOI |
10.1016/j.jbiosc.2017.02.017
|
PII |
S1389-1723(17)30004-X
|
PMID |
28319021
|
MeSH |
Animals
Antibodies / genetics*
Antibodies / immunology
Cytosol / metabolism*
Nucleoproteins / immunology
Peptide Library
Protein Engineering / methods*
Rabies virus
|
IF |
2.366
|
引用数 |
2
|
リソース情報 |
ヒト・動物細胞 |
Ba/F3(RCB0805) |