Nanobody screening

Single domain antibodies, also known as nanobodies, are revolutionary biopharmaceuticals due to their robust stability and strong binding affinity against their target proteins. Our laboratory creates nanobodies without any in vivo processes or sacrificing mammals, through several molecular biological experimental processes.

The libraries of three different kinds of scaffolds of nanobody are containing CDR3 region with high diversity. A strong binder selection cascade involving ribosome and phage display enables a thorough selection of the nanobody with strong affinity to its target molecule. A number of unwanted non-specific binders to the target molecule can result during immunization. With this in mind, the pure in vitro selection processes may overcome the drawbacks of immunization-derived antibodies.

In silico-based affinity maturation

In silico-based affinity maturation superior to traditional wet lab-based techniques, in terms of cost and labor. We generate prediction models of the selected nanobodies, and we use a variety of docking programs to virtually dock the nanobodies to their target molecule. We use nanobody-antigen complex models to validate their accuracy. Eventually, we can examine key amino acid residues to substitute to increase the nanobodies' affinity to their targets.

Screening platforms for Discovery of Molecular Interactions and Functional Disease Regulators

1. APEX2 proximity labeling screening
The engineered form of ascorbate peroxidase (APEX2) enables proximity labeling (PL) in living cells by using biotin phenoxyl radicals. By constructing APEX2 fusion anti-CD20 antibodies, we expect to analyze previously unidentified CD20-associated proteins that cooperate with direct cell death which can help in understanding antibody functions and mechanisms.

2. CRISPRi screening
CRISPRi technology can be harnessed to probe gene functions and identify novel targets for cancer immunotherapy. We are currently working on discovering the modulators of obinutuzumab-dependent direct cell death in B cell lymphoma.

Molecular pharming

Plants are considered attractive hosts for the mass production of recombinant protein drugs due to their economic efficiency, stability, and convenience. Successful production of antibodies in plant systems will not only lower the production costs, but also eliminate the risks from viruses in the mammalian host systems.

We are interested in establishing an alternative recombinant protein production system using a variety of plant species, such as Arabidopsis, Tobacco and Marchantia to develop future biopharmaceutical production platforms.