In the panorama of Chinese intellectual history, Mozi, the founder of the Mohist school in the pre-Qin era, may be regarded as the earliest pioneer of scientific thought in China. Drawing inspiration from the Mohist principle of "Promote the world's benefits, eliminate the world's harms", we affirm that the pursuit of technology must, above all, be directed toward the advancement of human welfare.
Inheriting the Mohist spirit, our Human Practices work pursues "How we influence the world and how the world influences us". Under the premise of serving human welfare, we interact with society, understand social needs, continuously engage with stakeholders, and thereby clarify our goals and directions.
Antibiotic abuse, identified by the WHO as one of the greatest threats to global health, food safety, and development, is a complex international problem that is difficult to eradicate. This requires us to comprehensively consider different stakeholders at various levels and stages in our Human Practices work. We should obtain external feedback through continuous interaction and promptly improve our research direction. For this purpose, we established a nested dual-cycle model of EFUV and DBTL. These two cycles are highly integrated and together constitute the core process of our integrated Human Practices.
In our project, Integrated Human Practices (iHP) is not an auxiliary component but a process that advances in parallel with experimental exploration and mutually shapes each other. Our project adopts a dual-cycle model:
Inner Cycle: DBTL Cycle (Design-Build-Test-Learn) focuses on experimental design, building expression systems, experimental testing and learning, thereby promoting optimization of LL-37 in expression, purification and function.
Outer Cycle: EFUV Cycle (Explore-Feedback-Update-Validate) explores social needs and ethical considerations, collects and integrates feedback from the public and stakeholders, updates project plans accordingly, and validates social acceptance and implementation feasibility — ensuring the project remains aligned with evolving expectations throughout its execution.
This dual-cycle nested structure ensures that technological progress and social feedback advance hand in hand, reinforcing each other.
Hover over any part of the cycle diagram on the left to view detailed information about that phase.
Through the integration of EFUV and DBTL, the project adopts a spiral circular structure, in which technology provides solutions for society, while societal needs, in turn, stimulate further technological iteration.
Clarifying which groups our project involves and their interests, expectations, and influence is key to ensuring the effectiveness of feedback and validation in the iHP cycle. Therefore, we conducted detailed classification of stakeholders involved in the project. Drawing inspiration from the Mendelow's Matrix, we analyzed their roles and influence in the project.
Stakeholder Analysis Matrix
The success of a biotechnology project depends not only on technical realization but also on its ability to withstand scrutiny in areas such as ethics, safety, equity, and social responsibility. Through systematic ethical analysis, the project seeks to ensure compliance with standards of fairness, safety, accountability, and social value, making the LL-37 project not only feasible but also justifiable.
Ethical Analysis Framework
In 2024, the World Health Organization again listed antimicrobial resistance (AMR) as one of the top ten global public health threats. A paper published in The Lancet warned that over the next 25 years, more than 39 million deaths globally could be directly related to bacterial resistance to antibiotics, with middle-aged and elderly people facing the highest risk. With the spread of antibiotic abuse, the emergence frequency of superbugs has significantly increased, and food safety and public health face unprecedented challenges. We realized that the resistance crisis cannot be solved by a single drug or technology but requires fundamental innovation in antimicrobial strategies.
For detailed information and sources, see the References.
To better understand solutions for antibiotic resistance, we first conducted extensive literature research. We also interviewed experts engaged in antibiotic-related research, hoping to gain insights into solutions for AMR. Our team visited Professor Xu Jing from the School of Food Science at Jiangnan University.
In our discussion, she emphasized the major obstacle in antibiotic governance - the diverse types of antibiotics make it difficult to find a standardized governance approach. This further confirmed our view that antibiotic problems can never be solved through simple governance alone, and continuing with traditional antibiotics is not feasible. Therefore, we urgently need to find substances that can effectively replace antibiotics.
Professor Xu Jing pointed out that multiple substances currently exist as antibiotic alternatives, with antimicrobial peptides (AMPS) being the most promising. After learning about this substance, our team decisively began exploring AMPS.
However, after extensive and extensive literature review, we found that current AMPS are mainly synthesized chemically at extremely high costs, making it difficult to achieve cost-effective replacement of antibiotics.
Interview with Professor Xu Jing on antibiotics and cutting-edge research.
Despite their broad prospects in replacing antibiotics, the high cost of chemical synthesis presents a major barrier to the widespread application of AMPs. Therefore, we wondered whether we could use synthetic biology methods for low-cost synthesis of AMPs. With this question, we found Professor Xia Shuqin from Jiangnan University. She has long been engaged in research on biological synthesis of short peptides, which are small, bioactive compounds ubiquitous in nature and predominantly characterized by their short length, amphipathic structure, and high cationic density.
Professor Xia showed considerable enthusiasm for our conceptual framework, noting the promising application of AMPs as antibiotic substitutes due to their novel mechanism of action. This insight, coupled with an evaluation of common antimicrobial peptide traits, directed our subsequent focus toward the human-derived peptide LL-37. Our selection of LL-37 was based on its identity as a naturally occurring human host defense peptide, which exhibits potent broad-spectrum antimicrobial activity and robust anti-biofilm capabilities. Notably, it demonstrates low minimum inhibitory concentrations (MICs ranging from 8 to 128 μg/mL) against a panel of clinically relevant drug-resistant bacteria, including Pseudomonas aeruginosa and Acinetobacter baumannii. The unique mechanism of action offers a therapeutic advantage by slowing the emergence of bacterial resistance.
However, natural LL-37 faces bottlenecks such as low expression efficiency, poor stability, and high production costs, severely limiting its clinical translation and application promotion. In 2023, multiple studies further pointed out that while the alternative potential of antimicrobial peptide drugs in the resistance crisis has been laboratory-validated, their industrialization still faces many challenges including expression system optimization, large-scale preparation, and in vivo stability.
Interview with Professor Xia Shuqin on insights and perspectives regarding antimicrobial peptides.
Subsequently, to engage a broader range of stakeholders and understand public perspectives on antibiotic abuse and the potential of AMPs as replacements, we conducted a targeted questionnaire survey. Within a one-month period, we collected 586 valid responses. Participants aged 18-25 accounted for approximately 63%, primarily students, while the remaining 37% included middle-aged and elderly respondents.
Analysis of the survey data revealed that respondents were most concerned about safety (approximately 75%), effectiveness (about 70%), and price (about 65%), with half (50%) emphasizing the necessity of official certification.
Regarding application scenarios, 65% of participants preferred uses in medical assistance or chronic care, and 25% supported applications in food preservation. In contrast, acceptance in acute medical scenarios was significantly lower. Overall, public sentiment suggests that LL-37 is perceived to have broad prospects; however, its successful implementation is seen to depend critically on cost control, rigorous efficacy validation, and the establishment of a compliant regulatory framework.
We also understood that to better clarify the importance of AMPs in replacing antibiotics, we inevitably needed to communicate with frontline medical workers. Therefore, we visited physicians at Jiangnan University Affiliated Hospital. Through exchanges with numerous doctors, we learned that antibiotic resistance not only significantly increases the risk of treatment failure but also substantially raises the social cost of healthcare, as superbugs caused by antibiotic resistance may prolong patients' illness duration while increasing medication burden. It's estimated that annual global direct medical expenditure due to drug-resistant bacterial infections exceeds hundreds of billions of dollars, further strengthening our determination to use AMPs to replace antibiotics.
Based on the above foundation, our goal became very clear: to use synthetic biology methods for efficient, high-yield, low-cost synthesis of LL-37. However, synthesizing a new substance in cells often faces many difficulties.
At the project's initial stage, following the advice of our primary PI, Chen Anqi, we initially selected four strains of Saccharomyces cerevisiae for preliminary testing. After using Western Blot (WB) for separation and identification of LL-37, we found that although the WB bands were bright, LL-37 could not be detected in the lysate supernatant, which was difficult to understand. To solve this problem, we contacted Dr. Sun Chixiang from Eli Lilly. Dr. Sun suggested we use fluorescence localization experiments to determine the location of LL-37 produced by cells. His solution for the WB experiment setback was insightful. After conducting fluorescence experiments, we discovered that LL-37 was adsorbed onto the cell membrane due to charge interactions. The successful localization of LL-37 greatly advanced our experiment. After successful localization, we designed a high-salt, low-pH extraction solution and successfully detected LL-37 in the lysate supernatant.
Consultation with Dr. Sun on wet-lab issues.
Encouraged by this progress, we set out to further improve the release of LL-37. Although high-salt extraction was somewhat effective, it remained incomplete. At this juncture, we sought practical advice from Professor Sun Shan from the School of Chemical Engineering, Jiangnan University, whose main research focuses on the application of nanomaterials in tumor therapy and wound healing. She suggested trying surfactant-assisted extraction, since surfactants can solubilize membranes and liberate membrane-bound proteins. Following their recommendation, we tested several mild surfactants at different concentrations. The results were striking — LL-37 release increased noticeably, yielding more material for purification.
Despite resolving the protein release issue, low expression levels emerged as our new bottleneck. To tackle this, we conducted comprehensive literature research and consulted Professor Li Jianghua, who specializes in yeast metabolic engineering. Notably, both sources indicated that gene knockouts targeting proteases or regulatory elements could enhance heterologous protein expression. Following this guidance, we constructed knockout strains and observed elevated LL-37 expression in several of them.
Another practical challenge was the cost of induction. Dependence on galactose for induction was prohibitively expensive for practical applications. Through literature review and discussions with Dr. Qu Tianzhi, our instructor, we learned that knocking out GAL80 could remove the dependency on galactose, enabling expression using more economical carbon sources like glucose. Therefore, We implemented this strategy and successfully achieved LL-37 expression under glucose conditions, dramatically reducing costs while maintaining efficiency.
Furthermore, purification became our most persistent obstacle. When purifying LL-37, we found that nickel column purification was remarkably inefficient. We subsequently tried multi-nickel column series purification, but the effect remained poor. Facing this obstacle, we consulted researchers at Tsingke, who pointed out that due to LL-37's low molecular weight, it might be difficult to bind to nickel columns. Additionally, during washing, high imidazole concentrations would wash away LL-37, but low imidazole concentrations would retain many impurities, making separation very difficult. They suggested two improvement methods: using magnetic bead purification instead, or semi-preparative liquid chromatography. After comprehensive consideration, we chose magnetic bead purification and found its efficiency significantly improved compared to nickel column purification.
While wet lab experiments progressed, our dry lab team developed the CytoFlow framework. To more comprehensively validate this framework's completeness and feasibility, we interviewed Professor Jim Chen from Massey University, an expert in computational biology.
He commented: "Your CytoFlow framework has a very innovative overall approach, especially applying reinforcement learning to peptide sequence optimization, which is a cutting-edge attempt in this field. However, I suggest strengthening structural considerations, such as incorporating AlphaFold's structure prediction results, to better understand relationships between sequence, structure, and function. Additionally, the model's interpretability could be improved - I recommend adding visualization tools to show key amino acid site contributions, which would be more user-friendly for biologists."
His suggestions made us realize the importance of structural information. Since AMPs like LL-37 only form α-helices, we focused more on sequence effects on function, but for proteins with more complex functions, their spatial structure cannot be ignored. Therefore, when subsequently improving our Cytopia cell factory, we will integrate structure prediction tools like AlphaFold3. For interpretability, we designed many comparative and ablation experiments to prove module effectiveness. Additionally, we created heatmaps that highlight amino acid sites contributing most to activity and provide biological explanations. These improvements will transform our system from a pure "black box" to a more transparent, trustworthy tool.
Consultation with Professor Chen on challenges encountered in dry-lab work.
For LL-37 variants designed through our computational models, we hoped to rapidly synthesize high-concentration variants. Kangma-Healthcode, founded by Dr. Guo Min and over ten returned PhDs, postdocs, and professors, is famous for D2P (cell-free protein synthesis technology). As a self-developed technology, D2P can custom synthesize proteins directly from DNA encoding. Compared to traditional cell culture methods, it can rapidly, high-throughput, and cost-effectively synthesize specific target proteins. After clarifying this, we quickly contacted staff Li Hongwei from Kangma-Healthcode. He indicated that we could indeed rapidly synthesize high concentrations of our computationally designed LL-37 variants through cell-free protein synthesis technology. Based on this, we tested using kits provided by Kangma-Healthcode and found bright WB bands, meaning LL-37 variants were synthesized at high concentrations. Therefore, we established deeper cooperation with Kangma-Healthcode and received their sponsorship of experimental resources.
Discussion with Kangma-Healthcode technical staff.
After months of experimentation, we successfully achieved preliminary preparation of LL-37. Although the product's yield and purity had not yet reached ideal standards, this was highly significant - it moved our project beyond pure theoretical derivation and model construction to having real "evidence" for validation. We regard this as a valuable opportunity to bring our result beyond the laboratory, actively inviting stakeholders from across society to examine our work and to share their perspectives and suggestions on LL-37, thereby further improving our project and assessing its market potential in the pharmaceutical industry. To this end, we have successively engaged with communities, hospitals, enterprises, academic institutions, and regulatory bodies, conducting a multi-level, multi-dimensional program of research and dialogue.
In community exchanges at He Yun Community, we mainly faced ordinary residents, including many young parents. We popularized the serious harm caused by antibiotics and introduced the advantages of antimicrobial peptide ll-37. They listened carefully to our science communication about LL-37, then raised numerous questions around practical needs. One mother mentioned she was very worried about her child's frequent exposure to antibiotics during growth, and antibiotic resistance made her anxious about her child's future health security; if LL-37 was truly a safe, natural antimicrobial solution, she would be very willing to try it. But then a resident immediately asked: "If its price is much higher than existing antibiotics, can ordinary families afford it?" These voices reminded us that public demand for new technology lies not only in novelty but also in affordability and accessibility. If the project ultimately cannot find balance between price and effectiveness, even with scientific potential, it will be difficult to truly enter family scenarios. This attitude of both expectation and practical consideration made us deeply realize that public desire and concern for new technology often coexist.
A college student on site asked: "This is made using synthetic biology technology, right? It still sounds a bit worrying. How do you prove it's truly safe and risk-free?" This question pointed out details we need to address - ordinary people naturally have unfamiliarity and concerns about professional technical terms like "synthetic biology" and "novel antimicrobial peptides." Even if we have laboratory-level safety data, to truly convince the public requires more accessible science communication and more intuitive safety verification process demonstrations to gradually eliminate the psychological barriers brought by this technical perception.
Additionally, A 65-year-old patient with hypertension and chronic bronchitis raised a question. She worried about: "The medicine prescribed by doctors gives me acid reflux, and I worry about interactions between blood pressure medicine, cough medicine, and other drugs. If your LL-37 can treat these conditions, is it oral or injection? Will it 'conflict' with my regular medications?" These words reflect the distinct particularities of elderly groups regarding medication. As a population with high chronic disease prevalence, they often require long-term multiple medications, making concerns about drug interactions particularly prominent. Due to limited mobility, they prefer convenient self-administered forms like oral or topical applications, generally showing psychological or practical resistance to injectable drugs. This reminds us that if we hope LL-37 covers broader user groups, especially the elderly, we cannot stop at single drug safety validation but need to conduct early interaction studies with common chronic disease medications. On the other hand, dosage form design should fully consider this group's usage habits and physiological characteristics, prioritizing development of convenient forms like oral or topical applications, avoiding elderly populations being deterred by inconvenient medication or potential drug conflicts.
Activities in the community
At Wuxi Traditional Chinese Medicine Hospital, we communicated with Dr. Ma Yaying from the Respiratory Department. As a frontline clinical worker, her thinking was both direct and practical.
First, she affirmed the clinical value of AMPs—finding antibiotic alternatives is particularly urgent as drug-resistant bacteria problems become increasingly severe. But she also frankly stated that clinical applications have extremely strict evidence requirements, needing not only complete toxicology data but also layer-by-layer verification through multi-stage clinical trials, from animal experiments to Phase I clinical trials verifying safety, then to Phase II/III clinical trials verifying efficacy and indications. Each step is governed by rigorous standards that must be strictly adhered to.
Additionally, she noted that the mild mode of action of AMPs constitutes a double-edged sword: while it is advantageous for reducing collateral damage to host tissues, it also underlies their primary limitation in acute infection scenarios. Such infections demand rapid and potent pathogen eradication, a requirement at odds with the relatively gradual efficacy of AMPs.
Consequently, the application requires precise positioning and should therefore be strategically focused on areas such as chronic wound management and topical anti-infective treatments, where conventional antibiotics are often limited or ineffective.
Dr. Ma also mentioned that antibiotics' role in acute infection treatment currently cannot be replaced, but clinical drug resistance has become a huge burden. She showed strong interest in AMPs but clearly stated: "Only when safety and cost are fully verified can antimicrobial peptides truly enter clinical practice. After all, when hospitals choose treatment plans, they must consider not only efficacy but also patients' economic capacity and long-term medication safety."
This feedback made us clearly realize that establishing medical value must be supported by complete experimental and evidence-based pathways, not just single laboratory data. Every step must align with actual clinical needs to avoid disconnection between technology and application.
Interview at Wuxi Traditional Chinese Medicine Hospital
During exchanges with Tsingke, researcher Wen Shishi mentioned that if LL-37 is planned for pharmaceutical use, it must strictly comply with GMP standards. This means small-scale laboratory procedures need to be transformed into fully traceable, standardized, quality-controlled industrial production processes. From raw material procurement to finished product inspection, every step must leave clear records to ensure product quality stability and consistency. These forms of feedback made us realize that, once applied in practice, the focus should shift from the question of "whether synthesis is possible" to "whether large-scale production is feasible." We also came to a deeper understanding that the core criteria by which the industry evaluates technological value lie in cost control and scalability. A breakthrough achieved in the laboratory attains genuine commercial value only when it can be translated into a scalable industrial solution.
The Marketing Department at China Resources Pharmaceutical Commercial Group Co., Ltd. offered a valuable perspective. They stated: "LL-37's technical direction is valuable, but two key problems must be solved: First, can it achieve scaled production? If production is unstable, no matter how good the technology, it cannot meet market demand. Second, compared to existing drugs, does it have competitive advantages in cost and effectiveness? If effects are similar but price is much higher, market acceptance will inevitably be affected."
They also referred to the case of Nisin, a well-established antimicrobial peptide product, emphasizing that its successful applications in both the pharmaceutical and food industries provide a valuable precedent for our commercial exploration. At the same time, however, they noted that Nisin exhibits clear limitations—for instance, its restricted inhibitory effect against Gram-negative bacteria—which precisely represents the point at which LL-37 may offer a significant advantage.
They reminded us that for LL-37 to smoothly enter the market, we must first clarify "in which scenario it has the greatest advantage." Market experts suggested we build an application priority map, gradually advancing from low-barrier fields (like medical devices, daily antimicrobials) to high-barrier fields (like medical wound care, anti-infection drugs), rather than initially targeting the most difficult medical replacement direction. This reduces early risks while accumulating experience and funding through low-barrier field applications. During the exchange, staff also asked: "The technology is indeed interesting and can solve many existing product pain points, but I'm most concerned about cost. Can you explain how LL-37's cost control compares to traditional antibiotics?" This question made us realize that although LL-37's biological synthesis cost has decreased compared to chemical synthesis, it's still high relative to the market. We took this question back to the laboratory, hoping to continue solving the cost issue through continuous improvement of biological methods.
Our interviews weren't limited domestically. In an interview with Professor Sabrina Tian, a microbiologist from Massey University in New Zealand, we gained a more international perspective. She noted that New Zealand initially imposed strict restrictions on the cultivation of genetically modified crops. However, with advances in science and societal development, the government gradually showed greater support, and modified fruits and vegetables began to be exported abroad, generating positive economic and social impacts. This suggests that, in the future, synthetic biology and genetic engineering are likely to gain wider acceptance and benefit a broader population. She also pointed out that current global research and application of AMPs is rapidly developing, especially as drug resistance becomes a global public health challenge. LL-37, with its natural antimicrobial mechanism, has broad application prospects. But she emphasized that different countries and regions have significantly different product classification standards for such bioactive molecules, requiring multi-stage strict approval including clinical trial data submission, pharmacological and toxicological evaluation, and production process verification, with approval standards and cycles varying greatly between countries.
Therefore, project teams must design differentiated development paths based on target markets. She also suggested that to advance LL-37 implementation, we must first clarify its specific medical application scenarios—whether for skin wound care or respiratory infection treatment, and what delivery form to use, such as gels, sprays, or oral formulations. Different application scenarios correspond to completely different R&D priorities and approval requirements.
Additionally, Professor Tian particularly reminded: "Different cultures may have very different acceptance of 'human-derived molecules.' For example, some Asian populations have high trust in 'human body-derived components,' while some Western populations focus more on whether they've undergone strict ethical review and safety verification. If you plan to enter international markets, this must be researched in advance to avoid product promotion obstacles due to cultural perception differences."
These suggestions made us realize that project prospects depend not only on "whether we can successfully develop" but also on "how to implement in different institutional and cultural environments," further strengthening our confidence in the project's future and prompting us to plan strategic layout with a longer-term vision, incorporating international market differential factors into early R&D and market research.
Interview with Professor Tian on antibiotic issues and antimicrobial peptides in New Zealand.
Finally, we visited Wuxi Market Supervision Bureau and had in-depth discussions with staff members Lu Peiyao and Zhang Mi. Unlike academic institutions and enterprises, their focus was mainly on regulatory compliance and approval processes, with every suggestion closely tied to "how to legally enter the market."
After understanding our project, Zhang Mi said: "LL-37 belongs to human-derived peptide molecules. Safety assessment of such components needs to be more cautious. From a regulatory perspective, we focus on two aspects: short-term use safety and potential risks under long-term consumption or contact scenarios. You need to provide more complete safety verification reports, including long-term toxicity experimental data, allergy test results, etc. These are core bases for approval."
Lu Peiyao added: "However, you don't need to worry too much. This type of technology to replace antibiotics aligns with current public health policy directions of 'reducing antibiotic abuse' and is encouraged at the policy level. If you can establish a complete safety assessment system and product lifecycle tracking mechanism—for example, traceability from production to sales at every step, with ability to quickly locate causes when problems arise—the likelihood of subsequent approval will be much greater."
They then explained in detail the general process for new biological products entering the market: First, complete systematic safety and stability verification to ensure product quality is stable within shelf life without safety hazards. Then, enterprises need to submit filing applications to regulatory departments. Application materials must include production process flowcharts, detailed toxicology experimental data, potential risk assessment reports and response plans. If involving pharmaceuticals, they must sequentially pass clinical trial approval, Phase II/III clinical trial data submission, drug regulatory department production site verification and registration approval. The entire process may take several years. They particularly reminded us that during product promotion, we must strictly comply with advertising laws and related regulations. We cannot describe LL-37 as a "universal antimicrobial agent" or exaggerate its efficacy, otherwise we might touch regulatory red lines during promotion, affecting subsequent approval. This exchange made us deeply realize that implementing novel AMPs is a long and arduous journey requiring multi-party cooperation and effort. Compliance is not an "add-on" in later project stages but permeates every corner of research, mass production, and promotion.
Discussion with Wuxi Market Supervision Bureau staff on relevant laws and regulations.
Therefore, all exchanges converged on a consensus: LL-37 has broad development prospects and can fully become an important supplement to antibiotics, creating value in multiple fields including medical, daily chemical, and food. But only through continuous breakthroughs in the two core issues of cost optimization and efficacy validation—both reducing production costs to market-acceptable ranges and proving actual value through multi-scenario validation—can it truly enter the market, integrate into society, and realize technology's ultimate value.
In exchanges with Dr. Ma Yaying and Professor Xie Yunfei from Jiangnan University, we discussed ethical dimensions involved in LL-37's practical applications. Their perspectives focused on real ethical issues that must be faced in technological development.
Dr. Ma pointed out that informed consent is not just a legal procedure but concerns patients' or users' true understanding. She emphasized that true respect lies in making information genuinely, completely, and equally accessible to everyone. "We cannot be satisfied with legally being 'informed,'" she said, "but must focus on whether the other party has 'understood.'"She illustrated this with an example: a label stating "contains human-derived components" could elicit rejection among certain cultural settings, while simultaneously serving as a mark of trust and efficacy in others.Therefore, their team collaborated with communication scholars from the project's beginning to design different versions of explanatory materials, even using community workers for face-to-face explanations. "Only this way does information become not cold clauses but a bridge for communication."
Regarding safety, Dr. Ma Yaying's attitude was particularly cautious. She admitted that molecules with immune activity like LL-37, despite broad prospects, are still like "a sharp knife" that must be properly controlled. Besides routine hemolysis and cytotoxicity tests, they suggested evaluating stability and activity changes in different body fluid environments like simulated intestinal and skin conditions. "We cannot skip these tedious but necessary steps just because we're eager to launch products," Professor Xie Yunfei added, "especially the lack of long-term tracking mechanisms, which will be the biggest ethical concern in the future."
At the public health level, Dr. Ma reminded us to be wary of the "technological optimism" trap. "LL-37 is not here to replace antibiotics," she clearly stated, "it's a partner, a supplement, not a savior." They hope to position such products as auxiliary solutions in specific contexts, such as chronic wound care or early intervention for drug-resistant bacteria, thereby avoiding public misunderstanding and subsequent abuse. More thought-provoking, they repeatedly mentioned accessibility issues—"If only a few people can afford it in the end, no matter how advanced, it hasn't realized public health value."
Subsequently, Dr. Ma turned the topic to cultural and belief aspects. She mentioned that in some cultures, synthetic biology or human-derived products might touch traditional taboos or religious doctrines. "This cannot be solved by saying science is fine," she said, "what we need is listening and adaptation." In promotional strategies, they also avoid using potentially controversial terms, instead adopting more neutral, local expressions.
We also interviewed Professor Zeng Xianghua from Jiangnan University Law School about legal and regulatory issues our project needs to address. He mentioned: Laws and compliance are like silent navigators. Intellectual property layout, data privacy protection, cross-border regulatory differences... these seemingly dry clauses are key to whether products can steadily enter different markets. He suggested introducing ethics and legal experts early to establish a complete responsibility mechanism from R&D to after-sales, including adverse reaction reporting, usage data tracking, and regular evaluation updates. "Compliance is not restriction but protection, an investment in innovation sustainability," Professor Zeng summarized.
Throughout the dialogue, a word he repeatedly mentioned: responsibility. It's not just about passing approval and successfully launching, but about continuous attention and commitment for the next five, ten, or even more years. Technology might iterate quickly, but ethics requires us to slow down, look back, and listen to voices that might be overlooked.
This exchange revealed a core consensus: ethics is not external rules to technology but a way of thinking inherent in the innovation process. It reminds us that true progress is not just about making something, but how to better integrate it into the human world.
Interview with Professor Xie on ethical dimensions of LL-37's practical applications.
A product's successful implementation requires not only technological breakthroughs but also clear market positioning and market receptiveness. Therefore, to ensure antimicrobial peptides' commercial feasibility, we interviewed experts from different fields for comprehensive assessment.
We interviewed Professor Teng Lefa, Vice President of the China Association of Higher Education Marketing Research, who focuses on theoretical research and practical applications in brand strategy, advertising, and international marketing. He provided extremely valuable opinions for our project's commercialization from brand marketing and business strategy perspectives. He pointed out that as a technology-oriented project, we should build a differentiated value system using our project's technological leadership as core value positioning. We need to note that for different application scenarios, we should create different application labels. For example, when replacing antibiotics in feed, we should emphasize "green farming compliance," targeting China's feed antibiotic ban policy with "antibiotic-free farming solutions," partnering with farming enterprises to create "zero antibiotic certification" demonstration bases. He emphasized that for our project to truly replace antibiotics, we must build brand trust. We should collaborate with authoritative institutions for clinical verification and ensure complete data transparency.
During the interview, Professor Teng showed great expectations for our project. He specified that after ensuring our project can truly replace antibiotics, we can achieve global market breakthrough in three stages, prioritizing target markets into three tiers. The first tier targets regions with severe antibiotic abuse like Southeast Asia and the Middle East. The second tier turns to high-end markets, obtaining BPR regulatory certification in the EU and ensuring FDA compliance in North America, entering high-end markets with medical-grade products. The third tier faces price-sensitive markets like Africa, promoting agricultural antimicrobial peptides, and if possible, attempting pilot projects with international organizations.
Professor Teng provided tremendous help for our project's commercial implementation and had a very long-term vision. We should actively seek relevant cooperation opportunities. Notably, Professor Teng pointed out that we must ensure these biosynthetic AMPs are protected by patent applications and comply with relevant laws and regulations.
To address Professor Teng's questions about intellectual property and legal regulations, we sought answers from legal professionals. For confidentiality reasons, we cannot disclose their identity here, so I'll refer to them as Lawyer Li in the following narrative. Lawyer Li stated that if we want to apply antibiotics to agricultural feed additives, we must apply for relevant qualifications for our products. Currently, according to Ministry of Agriculture and Rural Affairs Announcement No. 614, only Zhongnong Yingtai's Bacillus thirty-seven peptide has obtained relevant qualifications domestically. He emphasized we must ensure production process traceability, establishing a complete recording system from bacterial cultivation to finished products. According to his understanding, this should include fermentation parameters, purification steps, and microbiological testing reports for final products. After learning our product hopes to apply to the medical field, he reminded us that when designing clinical experiments, we can introduce independent data monitoring committees to reduce legal risks of subject rights damage. We must clarify precise classification in the medical field, not confusing medical device pathways with drug pathways.
Interview with Professor Teng Lefa
For intellectual property protection, Lawyer Li pointed out we should achieve global patent coverage, applying for patents not only in China but also entering Southeast Asian and Middle Eastern target markets through PCT routes. For production processes not yet patented, we need to sign confidentiality agreements with core technical personnel, referencing "Supreme People's Court Provisions on Several Issues Concerning Application of Law in Civil Cases of Trade Secret Infringement." He suggested we conduct quarterly electronic monitoring checks of confidential locations.
Professor Teng's Strategic Planning Discussion
After interviewing multiple experts, we gained detailed understanding of our project's commercial feasibility and clarified legal regulations we should reference and corresponding intellectual property protection. Our project shows great market potential and can propose our solution for antibiotic replacement. Below we analyzed our project's commercial feasibility from four dimensions.
SWOT Analysis of LL-37 Project
After achieving certain experimental progress, to better conduct integrated Human Practices activities, our team participated in the Jiangsu-Zhejiang-Shanghai iGEM Regional Meet-up in Suzhou on June 28. At the meeting, Dr. Bao Yuhan, PhD from Tsinghua University School of Public Management and iGEM Liaison Officer and Human Practices Committee Coordinator, reminded us to emphasize the narrative logic of HP stories, highlighting integration with experimental work rather than limiting activities to education. He also mentioned describing HP activity cycle logic, which has great practical significance for our integrated Human Practices activities. At this meeting, we also achieved deep exchanges and cooperation with numerous participating teams, laying a solid foundation for further cooperation.
On-site photos from the Jiangsu-Zhejiang-Shanghai iGEM Regional Meet-up
We also participated in the 12th Conference of China iGEMer Community & Synbiopunk 2025. During the roadshow session, team members Ren Yuxiao and Guo Zihao took the stage to introduce their research project in detail—efficient biosynthesis of LL-37. This project uses synthetic biology technology to achieve synthesis of human-derived antimicrobial peptide LL-37, with results widely applicable in pharmaceutical, cosmetic, and other industries. During three days of intensive exchange, our team maintained an open attitude, actively engaging in deep interaction with strong teams including Fudan and NJTech-China. From refining core technical details to potential optimization in experimental processes, to specific pathway planning from laboratory results to industrialization, both sides conducted multiple rounds of detailed discussion around these key issues.
During exchanges, team members not only shared breakthroughs and confusions in technical challenges but also exchanged experiences on practical issues like experimental data interpretation and solution adjustment strategies—such as how to improve target product expression efficiency, reduce scaled production costs, and balance technological innovation with market demands. This cross-school deep dialogue not only allowed our team to absorb advanced concepts from other teams but also provided new perspectives for exploration in synthetic biology applications, promoting teams' mutual advancement through collaboration. At 3:00 PM, the Synbiopunk Challenge award ceremony was held as scheduled. Awards were based on projects' scientific innovation, presentation impact, and social value. The organizing committee presented awards to outstanding presentation teams. Our team ranked 4th among 110 teams.
As an important exchange platform for iGEM events in China, this conference was not only a stage for university teams to showcase research results but also a link for academic newcomers to collide ideas and explore genetic engineering frontiers together. From laboratory innovative concepts to practical exploration of technological applications, over 100 teams brought their topics and enthusiasm for deep exchange, injecting new vitality into China's synthetic biology field development.
On-site photos from The 12th Conference of China iGEMer Community & Synbiopunk 2025
Based on current experimental data, through optimization of the expression system and purification process, we have preliminarily achieved a yield of approximately 68.7 milligrams of LL-37 per liter of fermentation broth, with an estimated unit production cost of approximately 3568 yuan per gram. Although this represents a significant reduction compared to chemical synthesis methods (approximately 10000 yuan per gram), there remains a certain gap from large-scale commercial application.
Moving forward, we will continue to focus on cost reduction and efficiency enhancement. Through advancing high-yield strain construction, fermentation process optimization, and purification workflow upgrades, we will further improve yield and purity while reducing overall costs, laying a solid economic and technical foundation for the industrial application of LL-37.
In the short term, given that LL-37 development still requires substantial funding (covering core aspects like sequence optimization, fermentation process development, and animal efficacy evaluation) and cost control breakthroughs, we plan to prioritize the pharmaceutical industry track: through establishing joint laboratories with innovative pharmaceutical companies, we'll advance preclinical research in phases (such as completing druggability evaluation and toxicology experiments), while simultaneously conducting small-scale clinical cooperation with medical institutions (such as emergency treatment exploration for multidrug-resistant bacterial infections), using a "clinical demand drives technological iteration" model to validate product value. Meanwhile, through industry forums, doctor-patient science communication salons, and other formats, we'll continuously convey the core value of "antimicrobial peptides' non-resistance advantage" to clinicians, patients, and the public, gradually transforming antimicrobial peptides from academic concepts to perceivable medical solutions, paving the way for subsequent industrialization.
As technology continues to mature, we will advance toward longer-term goals: pioneering functional short peptide industrialization and building a standardized "functional short peptide cell factory" platform. This platform will not only efficiently produce LL-37 but also adapt to different short peptide expression needs through modular design (such as anti-tumor short peptides, immune regulatory short peptides, etc.), achieving "one-click" customized production. Ultimately, we hope this platform will break the industry bottlenecks of short peptide drugs being "difficult to develop, expensive to produce, and narrow in application," promoting functional short peptides from niche research fields to large-scale medical applications, providing synthetic biology solutions for global health challenges like resistance crises and chronic disease management.
Wiki freeze, but our explorations never end.
Future Development Roadmap
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