Soon after becoming aware of the problems of mAb production under the trend of laboratory animal reduction, we contacted the National Laboratory Animal Center to confirm that the issue of experimental animals is not just discussed in the European region. We wondered whether there are relevant institutions that have responded to the trend of reducing the number of experimental animals. After searching for some information, we found National Laboratory Animal 3R Center in Taiwan and successfully contact Ph.D. Su and Ph.D. Chen from the center. After explaining our project to these two experts, they introduced us to current domestic animal-reduced experimental plans promoted and the trend of research. They affirmed our program and told us that our plan is what they anticipate the researchers in Taiwan do. They hope the researchers can try to figure out methods and designs to reduce animal use while achieving their purpose. They also gave us some suggestions about our wet lab experiments, such as selecting vectors to help us optimize our project. Finally, at their recommendation, we interviewed Zhu Zeng Hong, CEO of the Taiwan Animal Society Research Association, in the back-end implementation part of the project.

After accepting Prof. Chang’s suggestion, we used Lentivirus as a vector. Prof. Chang continued to pay attention to our project and provided some more advice. For instance, since we set our target users as biotechnology companies, it is crucial for them to lower the cost and achieve the most significant benefit. Therefore, Prof. Chang suggested we add some growth factors aimed at immune cells to better culture the cells. This way, we can not only improve the quality of hybridoma cells and mAbs but also implement our project more smoothly.

Apart from Prof. Chang, to examine our project's feasibility, we consulted Prof. Jan from the Institute of Medicine of Chung Shan Medical University for more advice. Prof. Jan specializes in microbiology and immunology, which correspond with our project. After proposing our project to him, he showed great interest in our project and was even willing to cooperate with us to help optimize the production of mAbs.

After we finished our preliminary design of the bioreactor, we presented our project and design of hardware to Prof. Tung from Academia Sinica, an expert in microfluidic systems. After understanding our design, he recognized our ideas and gave us extra suggestions on the bioreactor. First, he analyzed the advantages and disadvantages of the commonly used microfluidics for particle separation and cell retention while pointing out that there was still room for improvement for the spiral microfluidic we designed. He further suggested that we search for related references about tangential flow filtration (TFF) and try to figure out a design that can simultaneously tackle these two problems. Meanwhile, he assisted us in drawing the design layout and fabrication of the microfluidic chip. We followed his instruction, revised our design step by step, and finally created a product that satisfies our anticipation.

After confirming that our project is technologically feasible, we further invited our target user, a biotechnology company, for an interview. Fortunately, one of the professors in our school is the chairman of LEADGENE BIOMEDICAL Inc., a company equips with mAb producing service. We seized this opportunity and had several meetings with Mr. Zhuang to obtain some advice on technology and our value.

For the technological part, we initially aimed at making a bioreactor for hybridoma cells. However, as Mr. Zhuang mentioned, the Chinese hamster ovary cell(CHO cell) is the cell line in charge of the mass production of mAbs. If we can design a bioreactor for CHO cells, the value of our hardware will rise substantially. Therefore, we turned the target cells cultured into CHO cells and slightly adjusted the design to satisfy market demand better.

As for our value, Mr. Zhuang showed a positive attitude to our efforts in reducing laboratory animal use. Besides, since he considered it different between the opinions of the biotechnology industry and researchers and medical staff, we were suggested to interview these downstream customers further to evaluate the effectiveness of our project.

In the next stage of our Integrated Human Practice activities, we contacted Prof. Zhao with the assistance of NCHU_Taichung. She is a professor from NCHU and is an expert in hybridoma cells and mAbs production. After proposing our project to her, she showed a great interest in our technology and further asked for some details. During our meeting, Prof. Zhao mentioned that the most crucial factor of mAbs for researchers is the quality, namely the affinity. However, as she said, some mAbs from certain biotechnology companies are not qualified enough for their experiments. As a result, she considered it beneficial for researchers to a certain extent if our project succeeds.

Apart from the interview with a mAbs researcher, we met with the medical technologist Prof. Lee, who is also a professor of Pathology at our school. As an expert simultaneously engaging in research and clinical work, Prof. Lee gave us several suggestions about our project and spoke highly of our value. During the meeting, Prof. Lee said that the primary factors influencing the accuracy are the affinity and specificity of the mAbs. If these two characters were not good enough, the result of the test may turn out to be a failure. Therefore, Prof. Lee fully supported our project and the part of reducing laboratory animal use.

After several previous interviews, we determined that optimizing Hybridoma technology will benefit researchers. We also understood how biotech companies operate commercially and whether our plans are marketable. To confirm that our plan adjustment meets our initial expectations, we interviewed Zhu Zeng Hong , the Taiwan Animal Society Research Association's CEO, to share with him the implementation of our plan and possible future development. We were delighted to get his affirmation after sharing. He also provided relevant resources to make our program visible to more people. We also expected that we can make complete results to follow the international trend.

We believe that a significant research trend, such as reducing experimental animals, is worth noting for students, so we have introduced synthetic biology and practical animal-related topics in the middle and high school workshops. In the university sharing session, because the participants are all college students who will engage in biotechnology-related fields in the future, we shared more details of the plan and the development of laboratory animal reduction in our country. Through the questionnaire feedback, we believe that everyone has clearly understood the issue and what we plan to do after our sharing.