At the beginning of our preparation, we are fortunate enough to contact Prof. Hsieh from Tunghai university, who is also a technical advisor of GreAT Inc., a biotechnology company specializing in monoclonal antibody production. After we explained our preliminary ideas to him, he had great faith in our plan and gave us some advice on experimental design. Moreover, he also sponsored us by providing the original hybridoma cells producing antibodies against SARS-CoV-2.

After a discussion with Prof. Hsieh, we confirmed the direction of our project. However, we encountered difficulty when choosing the vector for our DNA sequence. Hybridoma is a cell line that simultaneously possesses the property of B cell, which can produce antibodies, and that of myeloma cell, which can keep on dividing without limitation of division number. If we use the usual lipofectamine method to transfect our construct, our target gene is likely to be kicked out after several times of division and causes severe problems for our project. In order to tackle this hardship, we consulted Prof. Chang, who has long been implementing transfection and cell experiments. During our discussion, Prof. Chang mentioned that Lentiviral vectors could successfully insert our target gene into the hybridoma genome and replicate with DNA in cells without being kicked out. Therefore, we decided to take Prof. Chang’s advice and use Lentivirus as our vector. Finally, our target genes were successfully transfected into hybridoma cells and express protein. For more information, please see the Proof of Concept page.

Taking the advice from Prof. Chang, we started to search for some references on the procedure of Lentivirus transfection. This process should have been time-consuming. Fortunately, our partner Mingdao introduced us to Ph. D. Chen, an expert in Lentivirus transfection, at the stage of brainstorming. We made good use of our spare time when we went to Mingdao and guided them with lab skills to talk with Ph. D. Chen. During our conversation, not only did Ph. D. Chen share his personal experience and tips with us, but he also provided us with directions for searching and adjusting the protocol from references. This meeting is hugely beneficial to our project, and it also helps us implement the subsequent experiments more smoothly.

Apart from the interviews regarding wet lab experiments, we also met several experts from the field of the dry lab. In iGEM competition modeling is an essential part of the verification of our wet lab experiments. As our primary PI, and also an expert in the field of modeling, Prof. Liu guided us to search for related references and uses the formula documented to establish our model. Meanwhile, when it comes to antibodies and antigens, the binding poses of two molecules are crucial information. Therefore, Prof. Liu also gave us preliminary knowledge of how to carry out homology modeling. He also taught us how to further use the results on antibody-antigen docking to lay a solid foundation for our project.

As for the section on hardware, we also consulted multiple professors for assistance. We hope to produce a biosensor of the bioreactor to monitor the quality of the antibodies produced. After reading some references, we narrowed our search to the self-assembled monolayer (SAM) biosensor. We then contacted Prof. Wang from NCHU. Prof. Wang is a professor specializing in researching SAM biosensors and electrochemical electrodes. He gave us several helpful technical suggestions and invited us to visit the Core Lab for Animal Embryo Engineering and Regenerative Biology of NCHU. These interviews helped us get acquainted with the production of SAM biosensors and stimulate multiple ideas about the design of our biosensor.

Aside from the biosensor, a bioreactor that can automatically sort cells and antibodies with a microfluidic system is our other goal. At the beginning of hardware designing, we thought it is too difficult to combine the biosensor and the microfluidic system, so we were not prone to complete the combination. However, after meeting Prof. Huang, an expert in microfluidic systems, he considered it possible for us to overcome the technical hardships with our resources and background knowledge. Therefore, we integrated these items into one hardware to unify every compartment and raised the value of every single item.