Human Practices

Yang Ding

Professor/(Distinguished) Researcher, School of Pharmacy, China Pharmaceutical University

Time: 2024.7.3

About Yang Ding

His main area of research is the process of disk-structured nanoparticles in drug formation and delivery, focusing mainly on their bridging with stem cells, and their application in drug delivery system of treating some diseases like tumor and central arteriosclerosis.

Why we talked to him?

First, our project partially overlapped with his area of research, we want to understand in more detail how the nanodisks are made and the feasibility of our project. Secondly, we would like to learn about the ADME process of nanodiscs in the human body, which will be helpful for our subsequent design of nanodiscs for drug delivery.

What we have learned?

• The significance of hydrogel drug delivery:Professor Ding Yang believes that the significance of hydrogel drug delivery is relatively small. Following her suggestion, we decided to remove the hydrogel drug delivery method from our approach.

• Importance of MSP's affinity and structure:The affinity and structure of MSP (Membrane Scaffold Protein) are crucial for supporting and forming the disc-shaped structure. In subsequent experiments, we will choose three robust covalent linkers combined with a flexible linker, Gslinker.

• Incubation process for nanodiscs: It was highlighted that during the incubation of nanodiscs, we should not mimic the preparation of liposomes. A gentler preparation method is needed, allowing ample time for the process. Ultrasonication and reconstitution methods should be avoided.

Hongliang Wang

Professor, School of Basic Medical Sciences, Xi'an Jiaotong University

Time: 2024.7.17

About Hongliang Wang

His research interests include modeling of viral infection, virus-host interaction, and antiviral drug development.The main research contents include antiviral drug screening and development, cloning and cell culture model construction of important medical viruses.

Why we talked to him?

In order to verify our ultimate antiviral effect, we must use some virus models, and Professor Wang is an expert in this area. In addition, we also want to understand the current status of some antiviral drugs, so that we can better understand the current market demand and find where we can break through.

What we have learned?

• Importance of transmembrane domains in proteins: Professor Wang emphasized that proteins capable of membrane fusion must have transmembrane domains, and it is preferable to use full-length proteins.

• Significance of CD4 glycosylation: The critical role of CD4 glycosylation was confirmed. Since yeast glycosylation differs from that of humans, it is recommended to use a eukaryotic expression system for producing CD4.

• Emphasis on innovative therapeutic concepts: Given the maturity of existing anti-HIV drugs, Professor Wang suggested that we focus on highlighting the novel therapeutic concept of intervening with enveloped viruses that directly undergo membrane fusion, rather than solely focusing on the relationship of the experiment to HIV treatment.

Zhanxin Wang

Professor, School of Life Sciences, Beijing Normal University

Time: 2024.7.22

About Zhanxin Wang

Why we talked to him?

When designing our nanodisks, we were considering using human intestinal membrane proteins but encountered issues with purification and yield. Additionally, we explored the possibility of using the CD4 protein as a simulation target but faced challenges in expression and purification. Professor Wang's experience in protein expression and nanotechnology made him a valuable resource for our project.

What we have learned?

• Challenges in protein expression and purification: Professor Wang highlighted the difficulties in expressing and purifying membrane proteins, including low yield and the potential contamination. He suggested exploring existing expression systems and considering protein synthesis as alternative approaches.

• Importance of specificity and selectivity: He emphasized the need for high specificity and selectivity in our protein design to ensure accurate binding to HIV and avoid unintended interactions. He discussed the potential of protein engineering to enhance binding affinity and specificity.

• Nanostructure utilization: We discussed the use of nanostructures like nanodisks and GFP tags to detect protein expression and assembly. Professor Wang provided insights into the feasibility and potential challenges of this approach.

• Innovation and feasibility: Professor Wang encouraged us to think creatively and consider novel approaches to increase protein-virus affinity, such as protein engineering and structure modifications. He also stressed the importance of considering experimental feasibility and potential challenges.

Yu Shang

Associate Professor, School of Life Sciences, Beijing Normal University

Time: 2024.7.22

About Yu Shang

Her research interests mainly focus on the core signal transduction and cellular metabolic regulation mechanisms of cells, especially in discovering important post transcriptional modifications of key signal proteins (including ubiquitination, methylation, etc.).

Why we talked to her?

Our experiment involves a large amount of protein design, purification, expression, and functional validation, and Yu Shang is an expert in this area. Therefore, we turned to her to get some advice on purification of membrane proteins and MSP. We also told her our concern about insufficient experimental time.

What we have learned?

• Differences between His tags and GST tags: GST tags can increase protein solubility, but they are relatively long and cannot be expressed if the protein is insoluble. His tags are relatively small and have little impact on protein structure. She suggested that we use His tags as the final purification handle.

• The role of a fusion partner: The addition of a fusion partner (male E) is necessary as it can assist in the proper folding of recombinant proteins, thereby increasing their solubility.

• The role of signal peptides: Signal peptides have functions such as directing, secreting, and participating in protein folding, ensuring that proteins can accurately locate, transport, and perform biological functions. Therefore, it is necessary to add signal peptides (IFNα2).

• Change to another virus: She said that if we chose membrane proteins of HIV to conduct our experiment, the cell expression was relatively. And she suggested that we could validate using another virus. So ultimately, we employed the VLP system to validate the effectiveness of our nanodisks.

Sen Li

Professor, School of Life Sciences, Beijing Normal University

Time: 2024.7.23

About Sen Li

He has extensive experience in the field of biochemistry and molecular biology. Innovatively, he introduced the concept of single-chain antibody molecular chaperones, identified and developed single-chain antibodies targeting several major disease-related proteins, and created a single-chain antibody-oligopeptide chaperone system that effectively aids in protein refolding. Additionally, he conducted a systematic study on how mixed crowded environments affect protein folding and aggregation.

Why we talked to him?

Our experiment involves a large amount of protein design and expression ,and Sen Li is an expert in this area. Therefore, we turned to him to get some advice on how to design our MSP. We also share our solutions about the problems on our experiment.

What we have learned?

• Foucus on MSP: To align with iGEM's focus on "engineering," it is essential for us to design and express proteins effectively. We recommend shifting our design focus to MSP .

• Change MSP linker: sGFP may not assemble effectively as an MSP linker. We suggest replacing it with a protein that has strong interaction properties. Ultimately, using high-specificity proteins capable of forming covalent bonds, such as spy/sdy/Snoop, as MSP linkers will be more effective.

• Use mCherry instead of GFP: Fluorescence can serve as a tool for characterizing protein interactions, and using different fluorescent proteins can help avoid interference between fluorescence signals. Therefore, we use mCherry instead of GFP in our multi-chain MSP designs to verify the successful cyclization of MSP.

• Design a system: To align with synthetic biology principles, we were recommended to design a system that regulates the strain's metabolism to switch between producing different MSP linkers. This idea inspired us to use recombinant enzymes to produce multi-chain MSPs, simplifying pathway design and experimental procedures, improving production efficiency, and enabling the controlled production of self-cyclized MSPs and multi-cyclized MSPs within the same engineered strain.

Li Tong

Associate Professor, School of Life Sciences, Beijing Normal University

Time: 2024.7.23

About Li Tong

Professor Li Tong has long been dedicated to studying the mechanisms and regulations of drugs. From a biochemical perspective, using molecular biology techniques and methods, her researches focus on the multifaceted effects of drugs on the body.

Why we talked to her?

Firstly, our product is supposed to be a therapeutic drug, and Professor Tong, who teaches Biopharmaceuticals, can provide us with some insights and supplements from the perspective of the drug design and production. In addition, we lacked experience in operating experiments independently and were uncertain about the experimental conditions of the school. We hoped that Professor Tong could give us some suggestions on experimental methods and design.

What we have learned?

• New methods for characterizing intermolecular interactions: In order to observe the particle size of the sample, the transmission electron microscopy and the negative staining method are used to select different dye solutions for substances with different electrical properties. The dynamic light scattering particle size of the sample is measured using a nanolaser particle size analyzer. Besides, circular dichroism is specifically used to observe protein structure. In the later stage, the protein can be separated and purified and sequenced with biochemical techniques.

• The differences between in vitro experimental results and in vivo actual effects: The physical and chemical properties in the body, as well as the number and activity of cells, make drugs much more complex in vivo than in vitro experiments. Also, interference should be taken into considerations. So we have to consider the actual effectiveness of the medication and the timing of administration.

Xiaoran Hao

Associate Professor, School of Life Sciences, Beijing Normal University

Time: 2024.8.28

About Xiaoran Hao

Her research focuses on the field of microbiology and serves as the technical advisor for the International Genetic Machine Engineering Competition (iGEM).

Why we talked to her?

Laboratory safety is the cornerstone of the iGEM project, and Xiaoran Hao has professional knowledge of laboratory safety. We asked her about the safety issues in this year's nanodisk project and the experience that others can learn from in our project.

What we have learned?

• Laboratory safety practices: Before conducting virus experiments, we need to confirm the virus is screened in the Catalogue of Pathogenic Microorganisms Transmitted by Humans. After that, we can carry out subsequent experimental work. At the same time, the viral proteins expressed in the experiments must be non-toxic and non-allergenic.

• GLP: Firstly, team members must wear lab coats when entering the laboratory, and strictly comply with the relevant national regulations when using corrosive and irritating hazardous chemicals. Secondly, carefully standardize the operation of equipment such as centrifuges, water baths, autoclaves, etc. Lastly, experiments can be operated in a biosafety cabinet when necessary.

• Experience accumulated during the project: She emphasized that when designing the experimental program, it was necessary to consider the existing conditions of the laboratory and do the experiments that could be done in the laboratory. In addition, the earlier the team members enter the experimental cycle, the better. This year, it took a long time in the concrete implementation of the experimental program due to the lack of practical experience.

• Clear our position: She also brought up a common misconception that when doing iGEM projects, iGEMers often try to make a new drug or material in a relatively short period of a few months. But for the competition, expressing the protein successfully and conducting a series of experiments to verify that it meets the desired vision is enough to achieve the gold medal. Her words make us even more determined to complete the project.

Chenxi Zhang

Founder of Xinjiang Yimoleculer Biotechnology Co., Ltd

Time: 2024.7.2

About Chenxi Zhang

Mr. Zhang is one of the founders of Xinjiang Yimoleculer Biotechnology Co., Ltd., located in Xinjiang Software Park. The company has independent molecular laboratory, cell laboratory, tissue culture laboratory, and is an innovative enterprise focusing on the research of animal and plant cell and molecular technology.

Why we talked to him?

He has extensive experience in the research and clinical translation of biological products, as well as the establishment and operation of biological enterprises, which can provide valuable suggestions for the industrialization of our project.

What we have learned?

• Persuasive experimental data: If we hope to turn our project into a product, we have to additionally solve following problems——What is the assembly efficiency of nanodisks? Does the entry of nanodisks into the human body causes rejection, and if so, what is the strength?

• Three steps to be on the market: After the project matures, animal experiments are required, which include modeling and simulation of pharmacology and functional verification of pathology. After passing the animal experiment, we will enter clinical trials. Then take the experimental results of the first two steps and discuss cooperation with investors.

• Guidance for research: Mr. Zhang further provided us with guidance on researches we were able to make at that time. In terms of epidemiological investigation, we can consult doctors from the infectious disease department of the hospital to obtain patient information and related data. We can refer to the data results released by the National Health Commission regarding the number of infections of infectious virus and cure rates. For enterprise research, he believed that we could start from targeted drug related pharmaceutical companies and participate in roadshows and seminars more often.

BioBAY

Suzhou Biomedical Industrial Park

Time: 2024.7.14

About BioBAY

BioBAY is a professional technology innovation carrier established by Suzhou Industrial Park for the development of the biopharmaceutical industry. There are three key industrial clusters: innovative drugs, high-end medical devices and in vitro diagnostics, and biotechnology.

Why we paid a visit to there?

We have had the idea to industrialize our project since we confirmed the topic of this year’s project. CAU-China sent us an invitation to BioBAY when we urgently needed experience from successful biomedical companies. Thanks to CAU-China, we learned a lot in BioBAY.

What we have learned?

• Project incubation process: It typically takes ten years for a project to launch the market after the laboratory validation. There are some stages we must undertake, such as drug evaluation, clinical experiment, IND new drug application NDA listing registration application and so on. Every stage is time-consuming but meaningful, so we need to be patient.

• Frontier biopharmaceutical field : We have come to understand the industrialization process of biopharmaceuticals, experienced the most advanced medical equipment currently available, and listened the development process of BioBAY from its inception to now.

Communication with Westlake-China

Time: 2024.7.18

About Westlake-China

Westlake-China earned nomination on Best Entrepreneurship Award in 2023 iGEM Competition. They are experienced in developing a product with real-world applications.

Why we talked with them?

This year is the first year for BNU-China to consider connecting our project with entrepreneurship. Lack in experience, we desperately need help from teams having successful experience. Therefore, we sent an invitation to Westlake-China to ask whether they could share with us. They joyfully agreed.

What we have learned?

• Entrepreneurial Framework: Westlake-China emphasized the importance of a structured framework for entrepreneurship, including market analysis, cost-benefit analysis, and understanding the target audience.

• Data Collection and Estimation: We need to do comprehensive data collection and the use of reasonable estimation techniques when direct data is unavailable.

• Cost-Benefit Analysis: Cost-Benefit Analysis is essential as well. They stressed the importance of considering all costs, including raw materials, equipment, packaging, and delivery, to ensure the project's economic viability.

• Project Impact: Moreover, to outstand the advantages of a project, we are expected to consider both the social and economic impact, and its our potential over existing solutions.

Jian Hou

Investor, BOHE Angel Fund

Time: 2024.8.3

About Jian Hou

He earned his PhD from the Max Planck Institute for Molecular Physiology in Germany. After graduation, he worked on the translation of fundamental research, and he is currently a partner at the BOHE Angel Fund.

Why we talked to him?

We would like to understand the advantages and shortcomings of our project from an investor's perspective. Additionally, we seek to learn about the necessary steps for commercializing the project.

What we have learned?

• Find the clinical needs: We need to identify the superiority of our treatment method in addressing clinical needs. For example, in the case of HIV treatment, if neutralizing antibodies can achieve the same or even better results, then developing such a system might not be necessary. Moreover, the system involves numerous components and a complex manufacturing process, which presents a challenge that needs to be addressed.

• The current state of innovative pharmaceutical companies: Firstly, innovative pharmaceutical companies face financial difficulties due to the market downturn, and the allocation of resources to such companies is not optimistic. Secondly, original innovation is challenging with long research cycles, high funding requirements, and regulatory approval issues. Lastly, bringing original innovations to clinical use and making them accessible to the public is a lengthy process. Therefore, innovative pharmaceutical companies need to identify their own strengths and leverage their uniqueness and originality.

• How to find our advantages: We should explore the advantages of our project from aspects such as efficacy, drug safety, and manufacturing processes.

The National Intellectual Property Center

Time: 2024.7.22

About the National Intellectual Property Center

The most authoritative intellectual property institution in China.

Why we paid a visit to there?

In terms of intellectual property protection, we had the opportunity to visit the National Intellectual Property Center and consult with IP experts to gain a comprehensive understanding of the steps, timeline, and materials required for patent applications.

What we have learned?

• Patent application process: Through discussions with the expert, we learned that the patent application process for innovative pharmaceutical technologies in China generally takes about three years, which includes the preparation of application materials, review, and final approval.

• Precautions for Patent Application in Our Project: For our nanodisc project, the decision on how to protect our intellectual property is critical, and requires careful analysis of our unique situation. Unlike traditional approaches where a single nanoparticle design might be patented, our technology allows for the customization of nanodiscs by altering their size and the membrane proteins they incorporate. This flexibility presents both an opportunity and a challenge in terms of intellectual property protection.

Hao Jiao

Professor, BNU Business School, Beijing Normal University

Time: 2024.9.2

About Hao Jiao

He is a famous economist. His research interests primarily focus on strategic management, entrepreneurship, innovation management, and corporate governance in the context of the digital economy.

Why we talked to him?

We want to get more information about how to start our business and what investors are most concerned about.

What we have learned?

• What investors are most concerned about: What is the target customer,and what are the market needs?

• Company positioning: Should we focus our company’s business on developing both nanodisc drugs and delivery systems, or should we concentrate solely on the development of nanodisc drugs? He suggested us to calculate the earnings. After estimating, we chose to develop both nanodisc drugs and delivery systems.

Yijia Li

Entrepreneur, Founder of the Cell Drug Translation Public Service Platform at the Tsinghua Yangtze River Delta Institute

Time: 2024.9.28

About Yijia Li

He is a scientist and entrepreneur with extensive experience in stem cell technology and biotechnology industrialization. In addition to his remarkable achievements in the field of stem cell technology research, he also collaborated with Academician Zhao Guoping to co-found the Shanghai Baoshan Synthetic Biology Translational Research Institute, which is dedicated to the industrialization and application of synthetic biology technology.

Why we talked to him?

Our project involves the application of nanodisc technology in the biomedical field, and Dr. Li's experience and insights in the industrialization of stem cell technology are crucial to our project. We would like to learn how to translate our nanodisc technology into actual medical products and would like to take this opportunity to inquire about relevant experiences in real-world operations.

What we have learned?

• Challenges and Opportunities for Industrialization: Dr. Li emphasized that industrialization of any technology or medicine is not an easy task and pointed out the lack of our market research, especially in the cutting-edge areas of science.

• Upstream and downstream construction: Dr. Li suggests that when we consider industrialization, we should not only focus on product development, but also consider how to construct a complete industrial chain. This includes a whole set of specific processes, which involve various links.

Conference of Beijing iGEMer Community

Time: 2024.6.29

About Conference of Beijing iGEMer Community

Since 2023, Conference of Beijing iGEMer Community has been held by BNU-China before CCiC. It is a nice opportunity for iGEMers in Beijing to communicate with each other and improve their project furthermore. This year, 8 teams participate in the conference.

Why we held Conference of Beijing iGEMer Community?

We are curious about other teams' fantastic ideas, and we wanna to get opinions on our project from other iGEMers. What’s more, following the similar process of CCiC, it is a platform for all teams to practicing introducing projects, showcasing highlights, exchanging ideas.

What we have learned?

• Clearly present the project: Given that nanodisks are a new concept to most people, we have known that, if we aim to make others fully understand our project, we need to expound the fundamental logic of nanodisguiser and present in a more attractive way in the limited time.

• New techniques used in the project: From projects of other teams, we have explored new knowledge that may be useful in our own project, such as OmpA, recombinase, Surface Display and so on. We then conducted brainstorms and optimize our project.

• Establish cooperative relationships: Thanks to this conference, we have stablish cooperative relationships with UCAS-China、BIT-China、CAU-China、Peking、PekingHSC、tsinghua-igem 2024、BUCT-iGEM. And we were keeping in touch with them throughout the entire iGEM process.

CCiC

Conference of China iGEMer Community

Time: 2024.7.11-14

About CCiC

CCiC is a national summit in the field of synthetic biology. It aims to provide a platform for the Chinese iGEM team to share resources, promote mutual learning and exchange. This year, it was held in XJTLU from July 15^th to July 19^th.

Why we joint CCiC?

It seems that it has become a tradition for every iGEM team in China to participate in it. What a nice opportunity to communicate with other teams!

What we have learned?

For experiments:

• Optimization of our verification system: The team who are working on VLP specially gives us the inspiration——why not we deliver our virus-like particles using this system? So we adjusted our verification methods later.

• Use of the recombinase: We communicated with the team focusing on the recombinase in detail, and designed a method to incorporate their system into our project, which was beneficial.

• Reconsider our design: We received many doubts on whether we could truly complete our project in such a short time, and the argument point was mainly on the expression of our membrane proteins. Ultimately, after much thought, we decided to put most of our efforts on the design of MSP.

For HP:

We are fortunate to talk with Ms. Zhang Nan, Global Vice President of iGEM Foundation Headquarters, who has lots of experience in HP works. And thanks to her, we got much more new ideas of the Human Practice.

• HP should be integrated throughout the entire process of your project: Whether it's brainstorming to come up with your project by identifying pain points in a certain area, absorbing opinions from all parties during the progress of your project to continuously optimize your project, getting feedback and evaluations from all parties to assess the value of your project, or comprehensively evaluating the strengths and weaknesses of the project when it is completed. ·How to highlight the value of homogeneous work: Your team needs to demonstrate why spreading this knowledge is necessary. Then, you have to prove that stakeholders have gained practical recognition and feedback on this knowledge, which further drives you to create a better curriculum. These are all important and must be clearly stated.

• The correlation between what HP does and what awards related to HP do: The two are not directly related. Awards related to HP are extensions and supplements to HP content, such as Inclusivity Award which does not require direct relevance to the project. But what HP does must revolve closely around the project.

• Avoid lengthy speeches: She said that when the judges mark our wiki, their feelings are the same as we hurry up with deadlines on final weeks. If they fail to get useful information after reading a few lines, for the length was particularly long and difficult to read, they may give up directly.