| RRC ID |
52142
|
| Author |
Nguyen TD, Takasuka H, Kaku Y, Inoue S, Nagamune T, Kawahara M.
|
| Title |
Engineering a growth sensor to select intracellular antibodies in the cytosol of mammalian cells.
|
| Journal |
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.
|
| Volume |
124(1)
|
| Pages |
125-132
|
| Published |
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
|
| Times Cited |
2
|
| Resource |
| Human and Animal Cells |
Ba/F3(RCB0805) |
