Research |
Synthetic 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 synthetic nanobodies (sybodies) without any in vivo processes or sacrificing mammals, through several molecular biological experimental processes. The libraries of three different kinds of scaffolds of synthetic nanobody are containing CDR3 region with high diversity. A strong binder selection cascade involving ribosome and phage display enables a thorough selection of the synthetic 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 sybodies, and we use a variety of docking programs to virtually dock the sybodies to their target molecule. We use sybody-antigen complex models to validate their accuracy. Eventually, we can examine key amino acid residues to substitute to increase the sybodies' affinity to their targets. |
Screening platforms to uncover the molecular mechanisms of anti-CD20 mAbs
Non-Hodgkin Lymphoma is a cancer generally developing in the lymphatic system. One of its therapeutics, obinutuzumab, is a humanized type II anti-CD20 monoclonal antibody (mAb) showing increased antibody-dependent cell-mediated cytotoxicity and induces direct cell death relative to type I anti-CD20 mAbs. However, molecular biological mechanisms and regulatory proteins related to induced direct cell death are poorly understood.
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. |