P09-04
De novo protein design of suitable binders for DNA origami-based devices
Jielin WANG1, Peiqi HUANG1, Hisashi TADAKUMA *1, 2
1School of Life Science and Technology, ShanghaiTech University
2Gene Editing Center, ShanghaiTech University
( * E-mail: tadakuma@iqb.u-tokyo.ac.jp )
DNA origami-based nanodevices integrated with protein are powerful tools. Multi-binder systems are a promising approach to capture, mark, and treat target molecules and cells. Using commercially available binders (antibodies), we have recently shown the potential of DNA origami-based multi-binder systems, where we have successfully captured exosomes in a size- and biomarker-specific manner. However, commercially available binders (e.g., antibodies and nanobodies) are mostly strong binders with high kon and low koff, while suitable kinetic characteristics highly depend on the target and aim. Moreover, to capture, mark, and treat multi-protein complexes, multiple binders recognizing specific target regions are required to ensure that each protein is recognized and targeted individually. Here, we try to overcome these challenges by designing binders using de novo protein design methods, attempting to obtain weak binders (high koff binders). This approach would allow each protein to readily dissociate due to the binders’ low affinity, while the entire complex could be captured by multiple binders. We will present our latest results regarding dry (design) and wet (biochemical experiments) approach.