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)