Human Practices

Overview

In our iGEM project, Human Practice (HP) activities were systematically integrated into both the design and experimental phases, ensuring our research remains relevant, responsible, and responsive to real-world needs.

During the Design Phase, we conducted community outreach activities—such as speeches and surveys at Erlong Road and Xinyuan Xili communities—to gauge public understanding and attitudes toward influenza and vaccination. These engagements revealed significant concerns about vaccine safety, low awareness of annual influenza mutation, and barriers to vaccination among the elderly. A public perception survey at Beijing XinAo Shopping Mall further highlighted that, while many are open to vaccination, the main factors influencing acceptance are safety, long-term protection, and ease of administration.

Building on these insights, we entered the Learn Phase by consulting experts from academia, industry, and Traditional Chinese Medicine (TCM). Industry interviews provided us with crucial technical advice for vaccine scale-up, formulation, and regulatory pathways. Academic experts deepened our understanding of the challenges in universal vaccine design, particularly the immunogenicity of conserved HA2 epitopes and the potential of novel delivery platforms like ferritin nanoparticles. Peer discussions with other iGEM teams helped us identify biosafety and dual-use risks and align with competition regulations.

By integrating these outcomes, we refined our project design to prioritize the development of a safe, broad-spectrum influenza vaccine using highly conserved HA2 epitopes displayed on ferritin nanoparticles. We enhanced our experimental protocols to include rigorous safety and immunogenicity testing, and we developed robust science communication strategies to address public concerns. This continuous feedback loop ensures our project not only advances scientific innovation but also addresses societal needs, regulatory standards, and public acceptance.

Design Phase Human Practice

Community Flu Awareness Speech at Erlong Road

Location: Erlong Road Community
Topic: The Importance of Influenza Prevention and Vaccines

On August 1, 2025, four members of PROFUTURE BJ：Shuya Yang, Feifan Hong, Motang Zhu and Qingyi Su made a speech about "The Prevention and Treatment of Influenza and the Importance of Vaccination" at Erlong Road Community. Before they gave the speech, the four members first conducted a survey in the community to gauge the public’s knowledge of influenza related basics, as well as their understanding of and attitudes toward vaccines and universal vaccines. They then further refined the speech content based on the conclusions drawn from the interviews.

During the speech, the team members explained the hazards of influenza, the differences between drug treatment and vaccine prevention, and the principles and benefits of vaccines. They also popularized knowledge about synthetic biology among the public. Finally, they emphasized the necessity and importance of universal vaccination and advocated for everyone to actively get vaccinated.

After the lecture, the Q&A session saw active participation from the audience. The primary concern raised was vaccine safety. Many attendees worried about potential side effects of vaccination and whether the vaccine had undergone sufficient laboratory and clinical tests. A common question asked was, "Will the vaccine be tested on mice before it is released?"

Three years ago, rumors spread online about COVID-19 vaccines, claiming issues such as "having side effects," "being ineffective," and "undergoing too few tests." These rumors led some members of the public to develop resistance to vaccines—a trend clearly reflected in the results of our preliminary survey. Among the 11 people who participated in the street survey, 5 explicitly stated that they would not get the influenza vaccine, citing distrust in its effectiveness, concerns about insufficient safety, and fears of side effects, thus expressing skepticism about vaccine safety.

Through this, the four members of the promotion team explained to the audience: "Vaccine safety has undergone rigorous testing, and all vaccines recommended by the community health service center for everyone to receive have obtained national approval. Therefore, you can feel at ease getting vaccinated."

Someone also asked, "Is the vaccine given to everyone the same every year?" This reflects that residents of Erlong Road Community do not have a thorough understanding of influenza viruses and vaccines. Among the 11 people who participated in the street survey, all stated that they had heard of the influenza virus. However, 7 respondents said they had only a "general understanding" or "little knowledge" of the hazards of the influenza virus, while 2 said they had no understanding of its hazards at all. When asked the question "Are you aware that the influenza virus mutates every year?", 5 people replied that they were "not sure" or "hearing about it for the first time."

Based on the questions raised during the presentation and data from street research, PROFUTURE BJ concludes that the necessity of vaccination and knowledge about influenza virus are not widely spread in the community. At the same time, during the interview, it was felt that there are more elderly people in the community, and they usually do not have companies to organize vaccination, so they cannot get vaccinated every flu season. This further highlights the importance of developing universal vaccines , which can enable vaccine recipients to develop antibodies against multiple viruses. The probability of infection by mutated viruses during influenza season will be reduced.

Autumn is coming soon, and we hope that PROFUTURE BJ's speech about influenza prevention and vaccine importance on Erlong Road can enhance the public's understanding of the influenza virus. And alleviate concerns about vaccination and value the role of vaccines. Actively receive the flu vaccine to prevent infection with the flu virus in this autumn and winter.

What We Learned

Our survey and community speech revealed significant gaps in public understanding of influenza, especially regarding annual virus mutation and the safety of vaccines. Many residents, particularly the elderly, were hesitant about vaccination due to concerns about side effects and insufficient trust in vaccine efficacy.

Integration into Project Design

These findings highlighted the urgent need for a universal influenza vaccine that reduces the burden of annual vaccination and addresses public safety concerns. Therefore, our project design prioritized the development of a broad-spectrum vaccine based on conserved fluval epitopes, aiming to provide cross-protection and minimize the need for frequent updates. We also decided to include clear science communication strategies and educational materials to address vaccine hesitancy.

Elderly Engagement on Influenza Vaccines at Xinyuan Xili

Location: Xinyuan Xili Community
Topic: The Importance of Influenza Prevention and Vaccines

Building on the findings from Erlong Road, the team next focused specifically on elderly groups at Xinyuan Xili, as the earlier survey showed that elderly people had significant concerns and hesitancy toward vaccination. This activity followed up on the identified barriers among seniors, aiming to clarify misconceptions and directly address their questions.

Date: 2025/8/7
Shuxuan Chen, Qingyi Chen , and Shuyu Yang from Profuture-BJ gave a speech about the importance of Influenza Prevention and Vaccines for these grandparents in the Xinyuan Xili Community.

They introduced the benefits and principles of vaccines to advocate for people to actively get vaccinated. And they also introduced some content about our project and knowledge of synthetic biology.

Before the speech, the speakers asked the grandparents if they were aware of the flu vaccine, and most of them said that they were not very familiar with its principles. They just had heard that it could prevent influenza, but they were not very willing to get vaccinated themselves because they believed that all medicines have some side effects and worried that the vaccine was bad for the body. Based on these viewpoints, the speakers explained to the grandparents that "Before the vaccine is released, it has undergone countless experiments and clinical trials. All vaccines available for inoculation on the market have been strictly reviewed and approved by the state. Everyone can be assured to get vaccinated without worrying about the vaccine being toxic."

After the presentation, these grandparents gained a deeper understanding of the principles of vaccines and vaccination, and raised some questions about vaccination, including "Does the vaccine have side effects?" "Is it necessary to get vaccinated if you don't go to crowded places?" "Can you get vaccinated if you have hypertension/diabetes and other diseases? Will there be adverse reactions from the medicine and the vaccine?" and similar questions. Speakers also sequentially answered their doubts and educated them on some precautions to be taken when getting vaccinated, such as "People with allergies to vaccine ingredients, those with severely weakened immune systems, pregnant women, and other special groups should not get vaccinated," "Stay in the hospital for observation for half an hour after vaccination, and only leave after confirming there are no allergic reactions or other adverse effects," and "Avoid eating spicy and stimulating foods after vaccination, maintain a healthy lifestyle, and get plenty of rest."

Some grandparents said that it is too complex to get different vaccines every year and worry that their bodies cannot handle it. Taking this opportunity, our speakers further explained that the project we are doing is based on this issue. Speakers explained to the grandparents in a simple way the role of the broad-spectrum vaccine in our project, which is to immunize against multiple flu viruses with a single vaccination. Most grandparents are very interested in our project, considering it a very innovative design. However, a small number of people are skeptical about the broad-spectrum vaccine. In response, the lecturers explained the principles of the vaccine and proposed the application of synthetic biology in the project. After gaining a deeper understanding, the grandparents expressed that they are willing to actively get vaccinated once the vaccine technology is confirmed to be mature.

On 28th July: Survey at Beijing XinAo Shopping Mall

On 28th July, The team member Qi Dong and Zhu Motang went to the BeiJing XinAo Shopping mall. We entered the mall at 12 noon for the questionnaire and until 6 pm, we ended the questionnaire. During the questionnaire, we roughly understood people’s understanding of vaccines, people’s vaccination in recent years, and people’s views on vaccines. And we have popularized people with relevant content about spectroscopic vaccines, including the benefits of vaccination, the advantages of spectroscopic vaccines, etc. In this process, we interview 63 people, and 35 people consider vaccination to be extremely important.Then we stored out 50 questionnaires among them, then we conduct analysis.

This is the question we interview:

Your age is?
What is your education level?
Your area belongs to?
Have you heard of the flu virus?
Do you understand the dangers of influenza viruses?
Do you know that the flu virus changes every year?
Have you or the people around you been affected by the flu virus?
Who do you think needs to get the flu vaccine the most?
Have you been vaccinated in the past year?
If you have not been vaccinated, what are the main reasons?
Are you willing to get a new vaccine with a longer lasting immunity?
Which one do you prefer to vaccinate the vaccine?

Then we used images to analyse, and here are the two most important images.

With these two icons as the core and other icons as the auxiliary, we came to the following conclusion: people are willing to get vaccinated while ensuring the safety of the vaccine, and are more willing to get spectral vaccines with a long-lasting immunity effect and a wide range of immunity. This coincides with our research direction, which shows that if our vaccine is successfully developed, there will be a good market.

What We Learned

We discovered that elderly populations are particularly concerned about vaccine safety, especially regarding chronic conditions (e.g., hypertension, diabetes) and the complexity of getting different vaccines each year. There is also a lack of understanding of modern vaccine principles.

Integration into Project Design

This feedback led us to design our vaccine delivery platform (e.g., ferritin vesica) to be as safe and broadly protective as possible, with clear contraindications and post-vaccination care instructions. It also encouraged us to explore vaccine forms (e.g., lyophilized powder) that are easier to transport and store, making annual vaccination campaigns more accessible for seniors.

Public Perception Survey at Beijing XinAo Shopping Mall

After engaging with both the general community and elderly groups, the team expanded outreach to a more diverse, urban population at Beijing XinAo Shopping Mall. The earlier activities revealed safety and awareness issues, so this survey was designed to quantify public attitudes, preferences, and acceptance of new vaccine technologies at a larger scale. This allowed the team to validate and generalize trends observed in previous community-specific engagements.

On the 1st of August, team members Shawn and Henry headed to Beijing XinAo Shopping Mall, a busy shopping mall situated at the center of the Beijing Olympic Park. From 1pm to 4pm, we interviewed a total of 27 people, mostly aged 26-40. The questionnaire focused mainly on three sections: people's understanding and awareness of influenza viruses, their personal experiences with influenza and vaccines, as well as their hopes and concerns for future technologies related to novel influenza vaccines. The goal of this questionnaire is to help us gain a better understanding of how people would react to a product like ours on the market, and make amendments to our project to better fit the market. It would also assist with the construction of educational presentations, as we would then be able to target areas lacking public awareness.

Below is the list of questions in our questionnaire:

What is your age? (<18 / 18-25 / 25-40 / 41-60 / 60+)
What is your education level? (High school or below / college or undergraduate / master's degree or above)
What type of city do you live in? (Top-tier / second-tier / town or village)
Have you heard of the flu virus? (Yes / no)
How familiar are you with the harmfulness of flu? (Very familiar / somewhat familiar / not very familiar / not familiar)
Before filling in this form, did you know that flu happens every year? (Yes / no)
Have you or someone close to you been affected by flu before? (Yes / no)
Who do you think needs to get the flu vaccine the most? (multiple choice: children; elderly people; people with chronic diseases; medical personnel (doctors, nurses, etc); normal adults; everyone)
Have you been vaccinated in the past year? (Yes / no)
If you have not been vaccinated, what are the main reasons? (multiple choice: there's no need to; worried about safety and side-effects of the vaccine; high prices; inconvenience due to location; lack of trust for the vaccine; other)
If there’s a new universal vaccine capable of covering more strains of flu for longer periods of time, would you go to get a shot? (Yes, gladly / could consider, based on actual situations / not sure, more information needed / not willing to get a shot)
Which method of vaccination is most acceptable for you? (oral / inhaled / injection)
If you have a concern for new vaccines, what is it? (multiple choice: safety and side-effects; lack of efficiency of the vaccine; too expensive; lack of long-term verification; distrust for new technology; other)

Survey Results

We discovered that out of the 27 people we interviewed, only one person had never heard of the flu before, suggesting good base public awareness of the situation.

Unfortunately, we discovered that nearly half of the interviewed people had not received a vaccine shot in the past year. This would mean a suboptimal vaccination rate, and insufficient protection against yearly flu.

Upon further questioning, we discovered that worry about safety and side-effects was actually the main reason for not taking a shot, though many still stated that they didn't feel the need for it, suggesting a lack of general knowledge on the importance of vaccines.

In relation to developing new vaccines and people's response to them, almost a quarter of the people stated that they would be happy to get a shot, and an additional 14 people replied that they could consider it based on the situation, which is just over half of the total number of people. This is promising for us, as it suggests that most people would be willing to try out new vaccines, and that our product would most likely perform quite well on the market if it is succesfully developed.

Additionally, oral and injection were shown to be the two most widely accepted methods of vaccination, which we could possibly implement for our project in the future. We would also have to make sure to provide people with more information on safety and side-effects of the vaccine, as over 90% of people stated that safety was a main concern for them when getting shots for new vaccines.

What We Learned

Survey results showed that willingness to receive new vaccines is largely dependent on safety and effectiveness. Public preference leaned toward long-lasting, broad-spectrum vaccines and convenient administration routes.

Integration into Project Design

We incorporated these preferences by focusing our efforts on the safety profiling of our vaccine candidates and exploring multiple delivery methods (oral, inhaled, injectable). We also refined our communication plan to proactively address concerns about novel vaccine technologies and side effects.

Learn Phase Human Practice

Discussion with Beijing Institute of Technology iGEM Team

Insights from public surveys and community engagement highlighted not only technical and social challenges, but also concerns that could impact project safety and compliance. The team then sought feedback from peers—such as the BIT iGEM team—to reflect on biosafety, dual-use, and regulatory issues, ensuring that the project’s direction would be robust and responsible.

On August 13, 2025, two Profruture-BJ team members, Shuxuan Chen and Shuya Yang, represented their team in a discussion with the Beijing Institute of Technology (BIT) team, led by Wen Junjie. During the discussion, both teams shared their project directions and progress. Through each team's introduction, we learned that their project focuses on oncology research. Afterward, we each raised and exchanged questions about the project and the competition.

I. Safety and Toxicity of the HA2 Fragment

1. HA2 Fragment Properties Assessment: Does the 16-amino acid fragment from the conserved HA2 stem region used in our project possess known virulence factor activities (such as cytotoxicity, membrane fusion activity, and pro-inflammatory properties)? Based on existing literature and our design (which consists of only 16 amino acids), how likely do you think it is to pose a significant biosafety risk?

The other party suggested that we search online for relevant information and regulations regarding this issue and then adjust our presentation based on official instructions. We could also email the official website to inquire about relevant issues and refine our design based on the competition rules.

II. Concerns and Mitigations Regarding "Dual Use" (How to Avoid Dual Application)

2. Specific Concerns Regarding "Dual Use": Regarding this project, is there a possibility that the immunogen could be inappropriately used to enhance viral production? How do you think this possibility can be mitigated?

Since our research primarily involves recombinant protein technology, it should not enhance viral production and should be safe for humans. Instead, we should be concerned that the recombinant protein may undergo proteolysis and change its shape after entering the human body, potentially preventing it from achieving the desired effect.

III. Regarding the Risks of Future Applications (Nanoparticle Delivery) (Are there any risks associated with using nanoparticles in the future?)

3. Will there be long-term retention or metabolic issues when using nanoparticles to deliver our recombinant proteins in the future?

When using nanoparticles for delivery, key considerations include the dosage and individual liver metabolism capacity. Each person's body's tolerance is different, so each case needs to be analyzed on a case-by-case basis.

Through this meeting, we gained a deeper understanding of the iGEM competition's rules, culture, and community. As a new team, this exchange not only helped us improve our project but also provided us with an opportunity to connect with other iGEM teams and the entire iGEM community.

What We Learned

The discussion helped us identify biosafety and “dual-use” risks, especially regarding the HA2 peptide and nanoparticle delivery. Peers emphasized the need for rigorous safety evaluation and regulatory compliance.

Integration into Project Design

We updated our future plans to include additional biosafety assessments (e.g., cytotoxicity and pro-inflammatory activity tests) and began planning regulatory consultations early. This will help mitigate risks and ensure our design aligns with competition and legal standards.

Expert Interview: Integrating Traditional Chinese Medicine with Flu Prevention

We are a student team participating in the iGEM competition. This year's project focuses on "influenza prevention and treatment". While reviewing a large amount of data, we also noticed that traditional Chinese medicine has unique insights and rich experience in influenza prevention and treatment. Therefore, we contacted Dr. Geng Xiaoxia, associate chief physician of Heji Traditional Chinese Medicine Hospital, for an interview to help us gain a more comprehensive understanding of the hazards and prevention of influenza. On August 13th, Wang Xintong from our team went to Heji Hospital for this interview. First, she introduced the background of the project, and then based on the project background, she asked the following questions.

How does traditional Chinese medicine (TCM) view the disease "influenza"? Is it similar to the commonly mentioned "cold damage" ("shanghan") and "warm disease" ("wenbing")?
In fact, they are similar. Previously, they were collectively referred to as pestilential diseases. Later, Western medicine renamed it influenza. However, influenza is mainly a term used in Western medicine, and the treatment methods differ greatly. In Western medicine, the main treatments are intravenous infusion for antiviral purposes, along with antibiotics. In TCM, treatment starts with antiviral therapy, using some Chinese herbal medicines.
From your experience, which groups of people with specific constitutions are more likely to "get hit" during the flu season?
Elderly people, children, and those with poor immunity are more prone to getting sick, while those with vigorous qi and blood and good physical condition have a certain degree of resistance.
What are the common methods of TCM in treating influenza? How effective are they? And does TCM treat based on the differentiation of individual constitutions?
It is certain that TCM treats based on the differentiation of individual constitutions. For people with poor immunity, in addition to taking Chinese herbal medicines, some external treatments may be added, such as cupping, bloodletting therapy, or acupuncture to expel toxins from the body.
Traditional Chinese medicine emphasizes "preventing disease before it occurs." How do you understand the significance of the saying "When healthy qi is stored internally, pathogenic factors cannot invade" during the flu season?
As we are approaching the transition from autumn to winter, allergic rhinitis may start to appear. Now is the time to begin prevention: enhancing immunity and regulating the spleen and stomach. This can be done either through Chinese herbal medicines or external treatment methods.
Traditional Chinese medicine has many methods to enhance resistance in daily life (such as moxibustion, dietary therapy, and daily routine regulation). Can these help people improve their resistance to influenza?
Yes, they can. In the Dog Days (sanfu days), people can use Tianjiu (a form of medicinal moxibustion using herbs applied to acupoints in hot weather), which includes exposing the stomach to the sun and "Dijiu" (ground moxibustion), all of which can enhance immunity.
Traditional Chinese medicine has the concept of "prevention first, prevention is more important than treatment" for epidemic diseases. Do you think some prevention and control methods in modern medicine (such as vaccines) also reflect this idea?
The role of vaccines cannot be denied. However, their effectiveness is premised on what is called "sufficient yang qi" in TCM. Many people feel that vaccines do not work because their internal yang qi is insufficient, which is closely related to their constitution.

Based on this interview, our team has gained crucial insights, which have shown us different treatment methods and perspectives regarding the influenza virus in traditional Chinese medicine. Although there are some differences between Chinese and Western medicine in terms of treatment, under Dr. Geng's explanation, we understand that they are complementary and can work together to solve problems. At the same time, we would like to thank Dr. Geng for his valuable suggestions.

What We Learned

TCM experts emphasized the importance of individualized prevention and holistic health in combating influenza, as well as the complementary roles of TCM and vaccines.

Integration into Project Design

We considered integrating TCM concepts into our public engagement and education, such as promoting synergy between vaccination and lifestyle-based prevention. For future iterations, we may also explore the compatibility of TCM adjuvants or immune modulators with our vaccine design.

Industry Perspective: Interview with Dr. Gao Feixia, Shanghai Institute of Biological Products

With increasing technical questions (e.g., about delivery forms, stability, and scale-up) arising from both public discussions and expert input, the team interviewed industry professionals to gain practical advice on production, safety, and regulatory pathways. This helped bridge the gap between laboratory research and real-world vaccine application

On 3rd. August, 2025, Stephanie Yang, Wendy Hong, Coco Wang, and Olivia Wang. These four students represented our team and had the honor to interview Dr. Gao Feixia from Shanghai Institute of Biological Products Co., Ltd. through online interview. Dr. Gao is the R&D Director, Associate Research Fellow, Postdoctoral Fellow in Bioengineering, Doctor of Medicine, Attending Physician, and a key member of the scientific research team at the Shanghai Institute of Biological Products Co., Ltd. She has a long history of experience in biological product R&D, focusing on the prevention of diseases such as shingles. She has participated in the development and clinical application of the shingles vaccine.

Production:

If our laboratory design is successful and we want to make this vaccine a product that can be used by many people, what do you think will be the biggest difficulty?
Successful laboratory testing only represents proof of concept; transforming these experimental results into a viable product requires subsequent experiments to verify translational feasibility. After confirming the vaccine's immunogenicity and protective properties, we need to scale up the antigen production process. To move from a small vial of recombinant protein in the laboratory to large-scale production using production equipment, we must utilize cells to express the protein. At this stage, we must pay close attention to the protein's expression level, binding efficiency, and stability. Furthermore, scale-up often introduces unwanted impurities, such as recombinant protein-expressing cells and residual DNA. Removing these impurities during large-scale production is significantly more challenging than in small-scale experiments. Finally, before a vaccine is approved for marketing, it must undergo rigorous testing and evaluation, including large animal studies and clinical trials. Clinical trials must involve a critical population size and undergo Phase II and III testing before they can be validated. This ensures the vaccine's effectiveness and safety, allowing for widespread adoption.
How do we know that the recombinant viral protein we produce is correct, pure, and safe? What basic tests are necessary?
First, regarding the accuracy of the recombinant viral protein, we need to ensure that its structure is correct. We can use instruments such as gel electrophoresis and mass spectrometry to confirm its structure. For example, if the desired protein structure is granular, we can use an electron microscope to verify that its primary and secondary structures are as desired. Scanning calorimetry and dynamic light scattering can also be used to determine the stability of the protein structure and whether its particle size and structure are correct. Second, regarding the purity of the recombinant viral protein, we can determine this by testing for residual protein residues and checking for residual DNA. Finally, we must minimize the endotoxin content in the protein before releasing it as a product. Third, regarding safety, since the vaccine is intended for human use, we must first ensure that it is sterile; any contamination should be avoided. Secondly, we need to test for abnormal toxicity. We can first administer it to animals to observe any adverse reactions to confirm its toxicity. Furthermore, we need to administer it to guinea pigs to ensure that the vaccine does not cause an allergic reaction. Finally, we can conduct multi-phase clinical trials using small-scale population experiments to observe whether the vaccine is safe.
If the immunogen is unstable after production, is there any simple way to protect it? (For example, adding a protective agent? Freezing?
If the produced immunogen is unstable, it will be difficult for us to proceed further, but we have certain ways to maintain the stability of the immunogen. First, for liquid dosage forms, we can use different buffer formulas or add some reagents to improve them. Second, we can add a protective agent to make it lyophilized. We can add some trehalose to the liquid dosage form and freeze-dry it in a freeze dryer to make it into a lyophilized powder. The lyophilized powder will have a longer shelf life.
Will the final universal vaccine be in the form of an injectable liquid? Or some other form (like a freeze-dried powder?) Which form is better?
This is based on whether our vaccine is stable. If it is not stable, we cannot store it in liquid form, so our better form is of course to make it into freeze-dried powder. The first aspect is that it is more stable. It can improve the stability of the protein and enable the antigen to work better. Secondly, freeze-dried products are easier to transport than liquid forms because they can be stored at room temperature for a period of time, so we can ship them to farther places, such as abroad. Moreover, the shelf life of freeze-dried products is longer than that of liquid dosage forms. However, the freeze-drying conditions also need to be optimized, so we need to decide according to the situation.
During the vaccine production process, how can companies ensure the quality of each batch of vaccine?
To ensure the quality of each vaccine batch, two key aspects must be considered. First, quality control must be performed during the production process. This means that the raw materials for each batch of protein are tested before they are expressed. For example, the culture medium and vector used to express these proteins are tested. These are only used after they have passed the tests. Second, during the production process, after the protein is expressed, tests are performed to check cell density, viability, and protein expression. Purity is also tested, such as for protein purity and impurities. Only after passing these tests will the company release the vaccine batch. Finally, ensuring high vaccine quality depends on the production process. To minimize variability between vaccine batches, companies also conduct verification of the production process.
When mass-producing vaccines in a factory, what are the main costs? (Equipment, labor, raw materials?) How do you estimate these costs? How can you minimize them?
There are three main aspects: equipment, labor, and raw materials. Large-scale production requires a large number of raw materials, and the combined prices of different raw materials can add up to a significant amount. Therefore, we control procurement costs. We estimate the total cost based on the amount of raw materials we will need. For equipment, we can't just estimate the purchase price; we also need to estimate its depreciation cost, amortizing its wear and tear over each year. The third aspect is labor. Labor costs account for a significant portion of the entire production process. When estimating labor costs, we need to calculate not only the cost of the workers but also the benefits they receive. Finally, there are the costs of testing. To ensure consistent quality across each batch of vaccines, we invest in testing. We also need to obtain compliance certification and construct a factory to ensure vaccine compliance. Building and maintaining the factory consumes a significant amount of capital, and we also need to estimate the utility and maintenance costs associated with its operation.
Are there any different production requirements for broad-spectrum influenza vaccines than for regular influenza vaccines?
There are significant differences in the production process between broad-spectrum and standard influenza vaccines. Standard influenza vaccines are also known as seasonal influenza vaccines. A key difference is that broad-spectrum influenza vaccines require greater flexibility and stability in their production processes. Seasonal influenza vaccines are formulated based on the strains prevalent each year, while broad-spectrum vaccines require no changes. They provide protection for many years without the need to change strains. Protein expression is relatively simple, as viral culture is less complex. However, the protein must be stable, ensuring that each batch is identical. Quality control is more stringent for broad-spectrum influenza vaccines, as standard influenza vaccines are only produced for one year and target a single strain, while broad-spectrum influenza vaccines are effective against a wide range of strains, requiring greater protection. Broad-spectrum influenza vaccines, which rely on a single HA2 target for weak immunity, require the addition of an adjuvant. Standard influenza vaccines generally do not use adjuvants, but to enhance the protective efficacy, immunogenicity, and cross-reactivity of broad-spectrum influenza vaccines, the requirements for adjuvants and adjuvant systems are more pronounced.
How can we be sure that this vaccine is a broad-spectrum vaccine, or can we never say for sure that it is a broad-spectrum vaccine, but it just has stronger immunity than ordinary vaccines?
We are currently unsure about the number of strains a vaccine provides protection against, and neither the US or European Medicines Agency (EMA) are clear. Currently, there are no clear standards for how many subtypes a broad-spectrum vaccine must cover, or what level of protection it must achieve to qualify as a broad-spectrum vaccine. We can only be certain if the approval agencies introduce relevant requirements in the future.

Research:

Besides HA2, what other targets or potential targets could be investigated?
HA2 primarily induces neutralizing antibodies. Neuraminidase (NA) is also relatively conserved, providing some cross-protection against viral spread. The conserved sequence of NA is a target. Secondly, internal proteins, such as NP and M1, are also relatively conserved. NP is also a relatively conserved matrix protein, while M1 primarily induces cellular immune responses, helping the body clear the virus. The matrix protein M2 envelope domain, also known as M2E, is also relatively conserved in influenza A and can also play a role in clearing the virus, but its immunogenicity is relatively low and requires interaction with other targets.
Is there potential for commercialization of our research results?
If so, what difficulties and challenges might we face? The primary issue is translational success, as we need to translate small laboratory batches into a larger population, which presents numerous challenges. The first is preclinical evaluation. We first test the vaccine in animals to determine its efficacy and safety. We also need to conduct large-scale human clinical trials, which present challenges and are time-consuming. Secondly, our broad-spectrum vaccine must ensure the longest possible duration of protection, a factor we must consider during clinical trials. Therefore, preclinical and clinical trials present significant challenges. Another challenge is defining immunogenicity and protective thresholds. We are uncertain about the specific strains we will target and the scope of protection, which also presents challenges. Fourthly, ensuring the stability of our production supply chain requires us to explore the possibility of scaling up the protein for large-scale production. Finally, regulatory approval is required from regulatory agencies such as the National Medical Product Administration (NMPA) and the Food and Drug Administration (FDA) before a vaccine can be successfully marketed.

Based on this interview, our iGEM team gained crucial insights that have greatly helped us in both the production and research aspects of our project. In production, Dr. Gao stressed that scaling lab results to mass production needs solving recombinant protein expression, binding efficiency and stability issues. Removing impurities like cells and residual DNA is far harder at scale. For quality control, structure verification uses gel electrophoresis and mass spectrometry. Purity control targets residual proteins, residual DNA and endotoxins. Safety validation includes sterility tests, animal toxicity tests, animal allergy tests and multi-phase clinical trials. Unstable immunogens can be fixed via optimization buffer or freeze-drying with trehalose added. If liquid dosage forms are unstable, freeze-dried ones are preferred because they have better stability and are easier to transport. Batch quality consistency requires quality control and process validation throughout production. Costs mainly come from raw materials, equipment depreciation, labor, testing costs and factory compliance costs. Compared to seasonal flu vaccines, broad-spectrum vaccines need more flexible production as there is no need to change strains every year. They also require stricter quality control standards and adjuvants to enhance immunogenicity. About research, Dr. Gao suggested alternatives to HA2: NA provides cross-protection, NP and M1 can induce cellular immunity to clear the virus, and M2E is conserved in influenza A. But M2E has low immunogenicity and needs to be used with other targets. She noted translation challenges: preclinical and clinical trials are time-consuming and need to verify long-term protective effects, the immunogenicity threshold is not clearly defined yet, mass production and expansion are difficult, and approval from the National Medical Products Administration and the Food and Drug Administration is required.

What We Learned

Dr. Gao provided technical and industrial insights on vaccine scale-up, purification, stability, and regulatory pathways. She stressed the importance of robust quality control, stability (favoring lyophilized vaccines), and the need for potent adjuvants in broad-spectrum vaccines.

Integration into Project Design

We refined our production workflow, adding steps for freeze-drying and adjuvant optimization. We also designed our experiments to compare ferritin’s ability to present short and long peptides, as this impacts immunogenicity and manufacturability.

Expert Interview: Influenza Vaccine Challenges with Prof. Deng

To address scientific challenges identified throughout HP activities—especially regarding universal vaccine design and immunogenicity—the team interviewed academic experts like Prof. Deng. Her input guided the selection of epitopes and delivery platforms, directly responding to both community needs (for a universal, safe vaccine) and technical hurdles mentioned in previous phases.

On the 31st of July, 2025, our team interviewed Professor Deng, PhD, senior researcher at the Chinese Academy of Sciences (CAS), to learn more about influenza vaccines and the potentials of our project. She is currently the head of the Molecular Virus and Vaccine Research and Development Team at CAS, focusing on research and development of influenza virus replication mechanisms and antiviral strategies. She earned a bachelor in biology from Lanzhou University and a PhD in Molecular Virology from Oxford University. Professionally, she has partaken in publishing 35 papers and 2 authorized national invention patents.

Through this interview, we sought to gain a complete picture about the past, present and prospect of influenza vaccines: what has been done, the challenges that they face, and current research directions, as well as an expert's opinion on our current research directions.

Q. What are the main types of flu vaccines currently available on the market? What characteristics and shortcomings do different vaccines have in terms of broad-spectrum efficacy and updating frequency?

Understanding the unique characteristics of influenza virus is crucial to understanding different types of vaccines. There are many types of viruses, including DNA and RNA viruses, which can be further divided into single-stranded, double-stranded, and circular forms. The influenza virus is a single-stranded negative-sense RNA virus, and there are 8 different gene segments for type A and type B influenza viruses. This special structure of its genetic material leads to the frequent mutations in the influenza virus... Therefore, due to those variations of influenza virus antigens, we need to continually research and develop new influenza vaccines annually. Currently, there are mainly four types of vaccines on the market: inactivated vaccines, recombinant protein vaccines, live attenuated vaccines, and mRNA vaccines—each with different trade-offs in immunity duration, breadth, and safety.

Q. When developing a universal influenza vaccine, which regions are usually considered first? What are your thoughts on the potential of HA2?
A. At the moment, HA2 is the most popular candidate antigen for universal influenza vaccines... However, there have not been any complete successes up to date.

Q. So what are the weaknesses of this solution?
A. The stem region is conserved but weakly immunogenic due to limited exposure; using it as an antigen leads to low antibody induction without additional engineering or boosting.

Q. Are there any current solutions for this, or is it still under research?
A. Strategies include cross-strain boosting and AI-guided design of exposed, immunogenic domains for recombinant expression and display.

Q. What are other prioritized targets besides HA2?
A. NA, NP, M1, and M2E offer complementary avenues (neutralization, cellular immunity, clearance) with varying immunogenicity.

Q. How is the acceptance of immunogen design based on structural biology in the development phase compared to traditional vaccine platforms?
A. Structure- and AI-guided immunogen design is valuable for discovering potent epitopes and enabling universal vaccines.

Q. What challenges do universities or student teams typically face when seeking to achieve industrial transformation of their early scientific research outcomes?
A. Preclinical model constraints, biosafety requirements (P2/P3), and long clinical timelines complicate translation.

Q. What is your opinion on ferritin?
A. Ferritin nanocages are promising and are advancing in clinical trials.

Q. If we don't have access to mice for testing, what alternative methods can we use to test our vaccine?
A. Virus-like particles (VLPs) can be used to test HA entry and immunogenicity in vitro.

Q. For the vaccines available on the market right now, how long does it usually take for them to go through clinical trials?
A. Typically more than ten years to obtain licenses and reach market.

What We Learned

Prof. Deng highlighted the scientific challenges of HA2-based universal vaccines, notably low immunogenicity and the need for innovative antigen design (e.g., AI-guided epitope selection, ferritin display), as well as the complexity of preclinical validation.

Integration into Project Design

We strengthened our focus on the structural and AI-guided design of HA2 epitopes and ferritin-based nanoparticle platforms. We also incorporated alternative validation models (e.g., VLPs) into our experimental plan, especially given potential resource limitations.

Expert Consultation: influenza Mutation Trends and Vaccine Evaluation with Prof. Zhu

To overcome critical scientific challenges in universal vaccine design and immunogenicity, we engaged with academic experts including Professor Zhu. His insights on epitope selection and delivery platforms proved instrumental in resolving both public expectations for a universal safe vaccine and previously identified technical difficulties.Professor Zhu's team researches the cellular mechanisms of phenotypic resistance in pathogenic bacteria and develops high-throughput single-cell analysis technologies. By integrating advanced imaging and sequencing approaches, the team systematically investigates phenotypic heterogeneity and evolutionary patterns of bacteria under antibiotic stress, providing novel theoretical insights and technical support for clinical anti-infective strategies.

I. Influenza Virus Mutation: Dynamic Evolution, Full of Challenges

Experts pointed out that influenza viruses, especially influenza A viruses, are characterized by rapid mutation. Every year, the surface proteins of the virus undergo gradual antigenic changes (drift), occasionally experiencing antigenic shift leading to pandemics. These dynamics complicate surveillance and vaccine strain prediction.

II. Vaccine Development: Facing Numerous Difficulties, Striving for Breakthroughs

The high mutation rate necessitates frequent updates and often short-lived protection. Seasonal vaccines are strain-specific; universal vaccines face technical hurdles due to diversity and manufacturing timelines.

III. Vaccine Evaluation: Strict Standards to Ensure Safety and Efficacy

Efficacy: neutralizing antibody titers, protection in challenge models, viral load reduction, durability of immunity. Safety: local/systemic reactions, immune-mediated pathology, long-term safety, and allergy/anaphylaxis monitoring. For ferritin and other nanoparticle platforms, consider innate activation, complement, scaffold-directed responses, and regulatory complexity; mitigate with surface engineering and rigorous preclinical profiling.

Expert Interview Transcript：

Key endpoints and evaluation strategies were discussed alongside structure/AI-driven immunogen design, ferritin platform advantages/limitations, and risks of non-specific immune responses with mitigation strategies (e.g., PEGylation, particle optimization, comprehensive toxicology and long-term surveillance).

What We Learned:

Experts outlined rigorous efficacy and safety endpoints for vaccine development, emphasizing the need for strong neutralizing antibody induction, broad protection, and comprehensive safety monitoring.

Integration into Project Design:

We integrated these endpoints into our animal and in vitro studies, ensuring our evaluation plan covers neutralization, protection, durability, and safety. These criteria will guide both our experimental design and future clinical translation efforts.