Human Practices
Antibiotic resistance is a major challenge in modern medicine that demands innovative therapeutic solutions. At the beginning of our project, we considered bacteriophages as an alternative treatment option against antibiotic-resistant pathogens. However, through continuous feedback from numerous interviews with experts in science, medicine, industry, regulation and ethics, we decided to shift our focus to endolysins, which are specific phage-encoded enzymes essential for bacterial lysis.
As the project progressed, we identified major needs in research of endolysins and developed a modular platform to efficiently identify and improve endolysins as potential drug candidates. Building on this, we combined the targeted use of endolysins and antimicrobial peptides (AMPs) to create an innovative strategy, especially against resistant Gram-negative bacteria. Additionally, the interviews revealed recurring concerns regarding the critical issue of biofilms in infectious diseases, implants, and food processing hygiene. Therefore, we adopted a synergistic approach by combining these specific proteins with bacteriophages to investigate enhanced methods for biofilm disruption and infiltration. Furthermore, we explored ways of future application possibilities in and on the human body to guarantee the actual feasibility of our approach.
To ensure that scientific innovations for antibiotic alternatives are not evaluated on their own, but fully embedded in medical, social, ethical, and regulatory contexts, we conducted a comprehensive stakeholder analysis. This included the use of a Power-Interest Matrix to strategically assess the influence and interest of different groups and involve them appropriately. Additionally, we performed a SWOT analysis to evaluate the strengths, weaknesses, opportunities, and threats of stakeholders, optimizing collaboration further.
This iterative development process forms the core of our Integrated Human Practices and is crucial for the sustainable direction of our project.
Antibiotic resistance is one of the greatest global challenges in healthcare today. Resistant bacteria complicate the treatment of infections, prolong illness, and causing an increasing number of deaths. As traditional antibiotics lose effectiveness and only a few new drugs enter the market, there is an urgent need for innovative alternatives (Clarici, 2025). Our project addresses this by exploring phage therapy and the targeted use of endolysins and antimicrobial peptides (AMPs) as novel approaches against resistant pathogens. Endolysins are enzymes produced by bacteriophages that degrade bacterial cell walls, while AMPs disrupt microbial membranes, leading to cell death ( Mba and Nweze, 2022; Idelevich and Becker, 2025).
In the medium term, our aim is to not only to contribute to academic research but also to provide industry partners with tools to efficiently identify new drug candidates. Ultimately, we envision the development of new therapies that physicians can use to treat resistant infections more effectively.
From the outset, we recognized that this endeavor raises not only scientific questions but also important societal, ethical, and regulatory issues. Therefore, we developed a structured approach to integrate these dimensions into our project. We combined our team’s expertise and interests to define a clear goal and design a plan for our Integrated Human Practices. The core was made up of fundamental values such as public health protection, transparent and fair handling of new therapies, and a commitment to making scientific results open, reproducible, and collaborative.
Our goal was also to develop sustainable solutions that preserve existing antibiotics and safeguard future treatment options. To put these principles into practice, we engaged with experts early on. Through a comprehensive stakeholder analysis, we identified key actors from medicine, academia, industry, ethics, legal and regulation along with their interests. The resulting interviews helped us understand diverse perspectives and actively incorporate them into our project design.
All findings are documented transparently under Integrated Human Practices, serving as a structured resource and as inspiration for future projects. In parallel, our educational and outreach activities raised awareness of antibiotic resistance across various societal groups, creating spaces for dialogue, reflection, and mutual learning.
To fully reflect the societal relevance and applicability of our project, it was essential for us to identify and include a diverse range of stakeholders in our Human Practices process. Rather than limiting our Human Practices work to technical aspects for the development of our modular platform or the use of phages and antimicrobial agents in a narrow sense, we were also eager to gain a deeper understanding of the overarching problem of antibiotic resistance and its scientific, societal, legal and regulatory dimensions.
Our project has points of contact with many different stakeholders, which we have grouped into four categories. We decided on the general stakeholder groups industry, academia, regulation & ethics and healthcare, from which we deliberately chose representatives to ensure diverse and comprehensive input.
Stakeholders from industry represent companies and professionals connected to the economic and technical aspects of our project. They provide valuable insights into market perspectives and potential applications. Engaging with industry helps us identify requirements for industrial implementation early on and align our project with real-world demands. There is growing interest in innovative antimicrobial strategies, highlighting the relevance of collaborating with industry partners.
Academic stakeholders such as researchers, PIs, and scientific experts contribute deep expertise, methodological skills, and the latest research findings. They play a central role in the scientific validation of our project and enhance its quality and depth trough critical reflection and theoretical grounding.
Actors from regulatory and ethical fields take on an evaluative and advisory role. They ensure that legal frameworks, safety standards, and ethical principles are adhered to throughout the project, which is why education is also important to us as it helps build public understanding and trust. Their perspective is essential for the long-term viability of new developments.
Healthcare stakeholders, including medical professionals and pharmacists, bring practical perspectives and experience. They help to identify potential hurdles and risks at an early stage and provide important insights into gaps in care and centered needs.
To strategically manage stakeholder engagement, we developed a Power-Interest Matrix (Figure 1) (Zhu et al., 2024). This matrix visualizes strategic overview of the influence and interest levels of different stakeholder groups in our project. The matrix is divided into four quadrants, which enable stakeholders to be addressed in a targeted and needs-based manner.
Figure 1: Stakeholder-Matrix.
In the High Power / Low Interest quadrant (A), we placed stakeholders with great influence but lower direct interest. This includes the Regulatory & Ethics group. Regulatory and ethical frameworks have a strong impact on the project, but their interest mainly focuses on safety, legal compliance, societal acceptance, and general developments in phage regulations, rather than the platform or biofilms specifically.
The High Power / High Interest quadrant (B) contains Academia and Industry. Academia holds high influence due to its research interests and methodological consulting, with strong interest in basic science, scientific collaboration, and recruitment of young scientists. Industry has moderate influence, as economic demands and market introduction will play a future role, but their interest is currently high because of the potential market and profit opportunities.
Healthcare stakeholders are in the Low Power / High Interest quadrant (D). They have strong interest driven by the urgent need for alternative therapies but have less direct influence. They primarily act as end users rather than decision-makers in development and implementation.
The Low Power / Low Interest quadrant (C) remained unoccupied, as no core project group fits this category.
This matrix helps us plan communication and engagement strategies precisely. Stakeholders with high influence and interest (Academia and Industry) are involved continuously and integrated into key decisions. Regulatory & Ethics, as powerful but less engaged stakeholders, are kept informed and consulted as needed to ensure compliance. Healthcare stakeholders are kept informed and included in feedback processes to better understand their needs.
To complement our Power-Interest Matrix, we conducted a SWOT analysis for each stakeholder group (Namugenyi, Nimmagadda and Reiners, 2019). This tool helps us evaluate the role and importance of stakeholders in our project and optimize collaboration. The four SWOT categories are defined as follows:
With an understanding of the SWOT terms, the assessment of the individual stakeholder groups could be carried out.
| Strengths | Weaknesses | Opportunities | Threats |
|---|---|---|---|
| Valuable market experience and practical insights into industrial implementation | Economic interests, internal processes and standards could prevent or slow down the integration of new technologies | Opportunities for valuable partnerships and practical adaptations | Competition pressure and risks from patents could negatively affect the project |
| Strengths | Weaknesses | Opportunities | Threats |
|---|---|---|---|
| Current expertise, methodological skills, and access to extensive scientific networks | Independent research interests can make it difficult to prioritize common project goals | Collaborations with various research groups allow access to new knowledge and innovative approaches | Possible conflicts between scientific priorities and project practicalities |
| Strengths | Weaknesses | Opportunities | Threats |
|---|---|---|---|
| Ensures adherence to legal and ethical standards, boosting societal acceptance | Regulatory requirements may reduce agility in decision-making and slow down innovation processes | Early engagement can ease approval and regulatory processes later on | Strict regulations and societal concerns could delay or complicate implementation |
| Strengths | Weaknesses | Opportunities | Threats |
|---|---|---|---|
| Practical insights into current therapies and needs for alternatives | Established clinical routines can hinder the introduction and acceptance of new procedures | Identifying concrete needs allows targeted development of new therapies | Challenges in integrating and accepting novel procedures risk project success |
When we first set out with our project idea, we quickly realized that listening to the people around us would be essential for guiding it in the right direction. Our Integrated Human Practices became a journey of exchange and collaboration. By conducting over 27 interviews with more than 40 stakeholders and experts from diverse fields, as well as engaging in discussions at the BFH MeetUp, we were able to systematically integrate different perspectives into our work and continuously refine our approach. Each conversation pushed us to look at our work from new angles, sometimes confirming our ideas, sometimes reshaping them entirely.
But our learning process did not end there. Through recurring discussions with experts, we established a continuous dialogue that accompanied us throughout the project. Their guidance did not simply answer our questions, it sparked new ones and provided crucial impulses, that made our project stronger and more meaningful at every step.
To present the findings from our interviews in a clear and comprehensible manner, we developed a uniform structure for documenting each conversation. Every interview was systematically documented according to four central categories:
The interviews were reviewed and discussed in regular meetings, ensuring that the insights gained were actively incorporated into the project development. In this way the voices of our stakeholders were not only heard but also effectively implemented.
Feel free to scroll through our timeline, click on the icons in the legend and timeline, and discover how our Integrated Human Practices approach has shaped and developed our project.
On March 17, an expert panel was convened to evaluate our early-stage project ideas. This interdisciplinary event brought together professionals from both the natural and social sciences, including renowned academics from our university. The panel provided valuable feedback that played a critical role in refining and reshaping our project direction.
The primary goal of the panel discussion was to obtain expert feedback on our project concepts from diverse academic fields. By engaging with experts in both the natural and social sciences, we aimed to identify potential scientific challenges, ethical concerns, and practical limitations at an early stage of development.
The discussion yielded several important insights that guided the evolution of our project:
The expert input significantly influenced the development of our project. Based on their recommendations, we made several adjustments:
The interview with Tim provided valuable insights into antibiotic prescribing behavior and resistance challenges from a pharmacy perspective. He described variations in how readily different doctors prescribe antibiotics and stressed the importance of cautious use to prevent widespread resistance. Therefore, he highlighted common clinical challenges, such as the frequent use of fosfomycin as a first-line antibiotic for urinary tract infections (short UTIs). Despite its convenience, this approach often results in incomplete efficacy and the need for subsequent antibiotic treatments. Furthermore, he explored the existing non-antibiotic alternatives and their limitations for serious infections. Thus, emphasizing the vital role that pharmacies can play in combating antibiotic resistance through education and stewardship. Tim also emphasized the importance of patient education. His observations underscored the impact of broader societal and agricultural factors on resistance.
We had the opportunity to discuss the extent of Tim's exposure to antibiotic resistance and how antibiotic prescribing patterns and patient practices manifest in the pharmacy setting. In order to ascertain the role of pharmacies and physicians with regards to the One Health concept we sought his views on communication gaps, collaboration with healthcare providers, and innovative approaches to antibiotic alternatives and stewardship.
Tim reported that many patients with UTIs often find the first prescribed antibiotic, typically fosfomycin, to be ineffective. Fosfomycin is commonly administered as a single oral dose and provides moderate eradication rates of 80–90%. However, it often does not fully clear the infection, necessitating follow-up treatments with other antibiotics. This pattern highlights the clinical limitations of current frontline antibiotic regimens.
Regarding alternatives to antibiotics, Tim noted that, although there are no direct substitutes for serious infections, phytopharmaceuticals (plant-based remedies) are sometimes used to treat mild UTIs or for prevention. However, antibiotics remain an essential component of treatment for more severe infections. This underscores the critical need to develop more effective alternative therapies.
Tim highlighted the importance of cautious antibiotic use and preventing resistance buildup. It is important to emphasize the significant role that agriculture plays in the dissemination of resistance. He stressed the need for rapid bacterial typing to enable targeted treatment and reduce unnecessary antibiotic exposure.
Financial disincentives limit pharmaceutical investment in developing new antibiotics. Tim advocates accelerating bacterial typing methods to enable faster, more targeted treatments, and he recognizes the potential of combining antibiotics with phages for future infection management. While pharmacies currently play a defined role in patient education and drug disposal, there is scope for expanded involvement.
Building on Tim’s insights, our project now prioritizes enhanced communication strategies aimed at healthcare providers and patients to raise awareness about antibiotic use. We plan to collaborate more closely with, for example, the elderly to develop educational awareness. Tim's observation regarding the restricted availability of alternative antibiotics prompts us to explore research opportunities in the field of phages and endolysins. Recognizing the critical role of agriculture in spreading resistance, we are strengthening our One Health approach by promoting the reduction of antibiotic usage in farming practices.
“Research into antibiotics should be funded by the state, as it is in the public interest.”
– Tim Barkow
Professor Dr. Ansgar Pommer supported and advised our project, offering his extensive academic expertise in molecular biomedicine, chemistry and biochemistry. Through his guidance, he clarified the complexity and urgency of the antibiotic resistance problem from an academic standpoint, strengthening our commitment to investigating alternative therapeutic strategies.
We engaged Prof. Dr. Pommer to obtain academic insight into the mechanistic and biomedical challenges posed by antibiotic resistance. We sought his perspective to validate and refine our research concept, as well as to reinforce the scientific rationale for focusing on alternatives to conventional antibiotics.
Prof. Dr. Pommer emphasized the multifaceted nature of antibiotic resistance as a global biomedical challenge demanding innovative solutions beyond traditional antibacterial drugs. Drawing on his background in protein chemistry and molecular biomedicine, he highlighted the limitations of current antibiotics. Because some antibiotics lose their efficiency against bacteria, for example when bacteria produce enzymes to destroy the beta-lactamase from penicillin. This is especially a problem in Gram-negative bacteria such as Escherichia coli. Thus, the need is rising to explore new agents, such as phages and endolysins. Furthermore, he emphasized that sustainable advancement in antimicrobial therapy requires rigorous scientific grounding, translational research, and the integration of interdisciplinary expertise. He also reaffirmed the importance of proactively addressing antibiotic resistance given its rising threat to global health.
Guided by the expertise of Prof. Dr. Pommer, we narrowed the focus of our project towards developing phage and endolysin therapies as promising alternatives to antibiotics. His view also emphasized the importance of basing our project on solid scientific frameworks while maintaining an interdisciplinary scope that bridges microbiology, molecular biomedicine, and clinical relevance. In That regard we planned to contact Prof. Dr. Axel Brakhage (an expert in the field of antibiotic-resistant pathogens) and Prof. Dr. Dobrindt (an expert in bacteria conjugation and phage research). This academic partnership has been instrumental in ensuring that our project advances with robustness, credibility, and purposeful impact.
To better understand the global challenge of antibiotic resistance, we interviewed Prof. Dr. Axel A. Brakhage, professor of microbiology at Friedrich Schiller University Jena and director of the Leibniz Institute for Natural Product Research and Infection Biology (Leibniz-HKI). Prof. Brakhage is an internationally recognized expert on pathogenic fungi, microbial natural products, and host-pathogen interactions. In our conversation, he stressed that the rapid spread of multidrug-resistant bacteria represents one of the most severe medical threats of our time. He highlighted both scientific and structural reasons for the crisis: resistance is an inevitable evolutionary consequence to antibiotic use, while the development of new drugs has stagnated for decades due to economic disincentives and political inertia.
Our goal was to gain expert insights into the mechanisms of underlying resistance development, the limitations of current pharmaceutical approaches, and the wider societal and economic context dimensions of antibiotic innovation. This enabled us to critically assess whether bacteriophage-derived endolysins can truly serve as viable alternative to antibiotics and to align his views with those of other experts we interviewed. At the same time, the discussion provided us with valuable confirmation of the scientific relevance and importance of our project’s focus.
During the interview, Prof. Brakhage explained that the rise of antibiotic resistance is not merely a consequence of misuse, but rather an inevitable outcome of evolutionary pressure. Any use of antibiotics, he emphasized, ultimately drives the emergence of resistant strains. At the same time, pharmaceutical innovation has stalled because large companies often consider antibiotic development economically unattractive. Short treatment durations, limited sales potential, and high costs of research and clinical testing make antibiotics far less profitable than drugs for chronic diseases, leading to a dangerous innovation gap. According to Prof. Brakhage, political frameworks and healthcare systems have likewise failed to establish incentives that could reverse this trend, and without systemic change, scientific discoveries alone will not be sufficient to overcome the crisis.
Drawing on his own research with the airborne fungal pathogen Aspergillus fumigatus, Prof. Brakhage illustrated how pathogens evolve sophisticated survival strategies. For instance, he and his team demonstrated that fungal spores possess pigments and surface proteins, that fungal spores possess pigments and surface proteins, such as DHN-melanin, which enable Aspergillus fumigatus to evade immune recognition and delay intracellular killing processes. These findings, he argued, exemplify the complexity of microbial adaptation, and highlighted the need for new therapies to be designed with a long-term perspective that anticipates such countermeasures. He further emphasized that microbes exist within ecological communities in which natural products, including antibiotics, play structuring roles. Novel antimicrobial strategies should therefore not be conceived solely as weapons to eliminate pathogens but as interventions that also take into account ecological balance and microbial interactions.
The interview supported our decision to focus on endolysins as an alternative to classical antibiotics. Prof. Brakhage’s remarks on the inevitability of resistance highlighted the importance of also considering combination strategies, such as coupling endolysins with conventional antibiotics or biofilm-disrupting proteins. His perspective further reminded us that the challenges of antimicrobial therapy are not only scientific but also economic and political, which we kept in mind when reflecting on the potential impact of our work.
“The future of multi-resistant germs is difficult, and I hardly believe that we can prevent it. We are talking about a silent pandemic.”
– Prof. Dr. Axel Brakhage
In South Africa, extreme social disparities reflect an underfunded healthcare system in which antibiotic resistance is widespread, but diagnosis and treatment are hindered by limited infrastructure and inconsistent guidelines. The overuse of antibiotics is further exacerbated by economic constraints, limited education, and lack of confidence in the healthcare system. The discussion highlighted the urgent need for cost-effective, user-friendly alternatives including endolysins and phage therapy, alongside improved education and consistent health regulations.
We interviewed Dr. de Beer to gain insights into the healthcare system and its challenges in South Africa, where antibiotic resistance is more prevalent than in Europe, and limited resources and access to healthcare increase treatment difficulties. The socioeconomic perspective of a country with huge inequalities and experience with public health interventions was of particular interest to us.
Dr. de Beer reported that 30 years ago, during her medical studies, viral infections and antibiotic resistance were already identified as emerging global health threats. She described South Africa as a country of contrasts in its healthcare system: while the wealthy receive excellent care, the poor are often underserved due to chronic underfunding. Antibiotic resistance is a growing problem, with resistant cases frequently observed in common diseases such as urinary tract and respiratory infections. Due to limited infrastructure, antibiograms are usually available only to very wealthy patients in town hospitals; for others, physicians must try the next available antibiotic when treatment fails.
Dr. de Beer highlighted the absence of consensus among healthcare providers: although physicians are formally required to attend training courses and conferences, strict adherence to treatment guidelines is rarely enforced, resulting in discrepancies in prescribing practices. Economic pressures exacerbate misuse: uninformed patients often demand antibiotics, even for viral infections, and may switch doctors if demands are not met. Many discontinue treatment as soon as symptoms improve, particularly in rural areas where follow-up prescriptions are difficult and taking time off work to reach a clinic is often impossible.
Additional challenges include low hygiene standards and limited public trust in the healthcare system, rooted in insufficient education, which she identified as the biggest obstacle to introducing new treatments. According to Dr. de Beer, any viable alternative to antibiotics must be affordable, easy to administer by well-trained personnel, accessible in remote regions, and ideally storable without refrigeration or strict sterility requirements – “something that can be used even in the middle of nowhere.”
The discussion highlighted the challenges of treating particularly vulnerable patients with antibiotic-resistant strains in an underfunded healthcare system. It underscored the urgent need for comprehensive healthcare strategies with consistent and enforceable treatment guidelines. At the same time, it reinforced our commitment to strengthen education and public awareness while advancing the cost-effective production of endolysins, which could represent a realistic and accessible therapeutic alternative.
“30 years ago, I was told that viruses and antimicrobial resistance are problematic emerging health threats and will play a big role in the future, and here we are."
“Thank you, guys for doing such an important job, we need an alternative solution urgently.”
– Dr. Ingrid de Beer
In South Africa, pharmaceutical research faces significant limitations due to underfunding and a predominant focus on high-mortality diseases, while antibiotic resistance has so far lacked strategic prioritization. Affordable prices, economic relevance, and effective marketing are crucial for alternative therapies to succeed, as many people prefer traditional healing practices and there is a lack of education. At the same time, relatively low regulatory barriers present opportunities to establish innovative approaches such as phage or endolysin therapy if given adequate funding.
We interviewed Ms. Lichtenberg to obtain insights into pharmaceutical research in South Africa, where antibiotic resistance already represents a very urgent challenge. Especially multi-resistant infections are more widespread there than in Europe, and in combination with limited access to healthcare and limited resources, South African researchers must deal with even more difficult circumstances. We aimed to identify the needs for treatments to improve the applicability of our concept.
Ms. Lichtenberg explained that research in South Africa is severely constrained by limited government support and chronic underfunding of the healthcare system. Current efforts focus primarily on urgent challenges such as HIV and tuberculosis, while comprehensive strategies to address antibiotic resistance remain absent. Nevertheless, a novel disinfectant called ‘Trifectiv’ was developed at Stellenbosch University while researching compounds effective against bacteria.
For any antibiotic alternative to succeed, it must be accessible to economically disadvantaged populations while remaining economically viable for local pharmaceutical companies. Major obstacles include limited public education and widespread reliance on traditional healing methods, witch doctors, or information from social media rather than pharmaceutical experts. Effective marketing and education are therefore essential to foster acceptance and build understanding of therapeutic mechanisms. Greater public awareness could also stimulate increased government funding.
On a positive note, South Africa has comparatively few regulatory restrictions on research. With sufficient funding and targeted communication, innovative therapies such as phage or endolysin treatments could be successfully established.
The interview highlighted that the success of innovative therapies depends not only on science, but also on effective marketing and targeted education, especially in communities with little prior knowledge. Scientific progress alone is not enough; it must be clearly communicated to create understanding and acceptance. With this in mind, we are dedicating efforts to improving our public relations work to make complex scientific topics more accessible and comprehensible.
Ms. Mavrina gave us insights into the responsibilities of hygiene officers, which include staff training, coordinating isolation protocols, and maintaining communication with the hospital hygiene department. She emphasized that routine MRSA screening is mandatory for new patients, particularly those from risk groups, such as farmers or other patients with a history of MRSA infection, to ensure early detection of resistant bacteria. She also highlighted that the correct use of protective equipment and strict hand hygiene remain the most decisive factors for preventing transmission.
Regarding antibiotics, she observed that therapies are increasingly targeted and culture-based, reducing unnecessary prescriptions. At the same time, she noted a concerning rise in multidrug-resistant organisms, underlining the need for cautious and responsible use of antibiotics.
On novel therapies, Ms. Mavrina was aware of antiseptic and antibiotic practices but had limited knowledge of phage therapy. She expressed some caution but showed openness to the idea, provided that clinical studies demonstrate safety and efficacy.
We contacted Anna Mavrina, a nurse and hygiene officer at a German hospital, to gain first-hand insights into the handling of multidrug-resistant organisms (MDROs) such as MRSA, VRE, and Clostridioides difficile in the clinical setting. Her dual role as caregiver and hygiene representative allowed us to understand both the practical application of infection control measures and the broader challenges of antibiotic resistance.
Multi-resistant pathogens constitute a persistent and growing challenge in modern hospitals. Their presence is not limited to isolated cases but increasingly affects everyday clinical routines, and some healthcare professionals perceive their incidence to be on the rise. This development underscores the urgent need for effective infection control strategies that extend beyond ad hoc responses and become firmly integrated into the structural processes of clinical care.
A cornerstone of such preventive measures is the consistent application of screening protocols, which enable the early identification of carriers or infected patients. In combination with targeted isolation procedures, these measures form the first line of defense against nosocomial outbreaks. Central to their effectiveness are hygiene officers, who function as crucial intermediaries between regulatory guidelines—particularly those issued by the Kommision für Krankenhaushygiene und Infektionsprävention (KRINKO) and the Robert Koch Institute (RKI)—and the realities of daily clinical practice. They not only communicate recommendations but also ensure their practical translation into workflows, routines, and staff training.
Another key pillar in addressing antimicrobial resistance is antibiotic stewardship. Rather than relying on broad-spectrum antibiotics as a default strategy, therapeutic decisions are increasingly based on laboratory diagnostics. The use of culture confirmation and antibiograms allows for targeted interventions, thereby reducing unnecessary antibiotic exposure and slowing the development of further resistance.
Despite these established practices, innovation in the field of anti-infective treatment remains limited at the clinical frontline. Novel approaches such as bacteriophage therapy, although discussed in scientific and translational research contexts, are still largely unknown among everyday hospital staff. Nevertheless, there is a cautious openness toward such alternatives, provided that their safety and efficacy can be convincingly demonstrated in clinical trials.
The interview showed us that strict hygiene protocols and targeted antibiotic use are essential pillars in controlling resistant pathogens. This confirmed that our project should not only consider innovative treatments such as phages but also align with existing hygiene structures in hospitals. The cautious, but open, stance towards phages revealed that clinical acceptance depends strongly on clear evidence of safety and effectiveness. Additionally, her concerns about phages potentially persisting in the body highlighted the importance of transparent risk communication and broad education among various groups of people in our project.
“It seems to me that multidrug-resistant bacteria are on the rise and are also impacting younger individuals.”
— Anna Mavrina
In conversation with Karin Moelling, a renowned cancer researcher, virologist, and expert in phage therapy, we gained valuable insights into the long history of research into phage therapy. She emphasised the growing importance of phages considering increasing antibiotic resistance, but pointed to existing public scepticism and strict legal and costly production requirements in the European Union and Germany. At the same time, she recommended many other researchers and experts who are working with different perspectives on phages.
Karin Moelling was the first professor we consulted on phages. She is a renowned virologist, author and expert in the field of phage therapy. Furthermore, she initiated with others the Phage Germany project (www.phage Germany) and is engaged in overcoming societal distrust toward phage therapy. Our aim was to obtain her expert assessment of our project, to deepen our understanding of phage therapy, and to gather her perspective on the escalating threat of antibiotic resistance.
Professor Moelling explained that phage therapy has been practiced in Eastern Europe, especially during recent wars, for more than a century, particularly at the Eliava Institute in Tbilisi, Georgia. She emphasized the growing threat of epidemics as well as the critical importance of addressing antibiotic resistance and highlighted the potential role of viruses, especially bacteriophages, at least transiently, until we have new antibiotics. While welcoming our commitment, she reminded us that several companies are already pursuing similar approaches and referred us to numerous scientists and firms working with bacteriophages who could serve as valuable contacts.
She explained that although phages are produced in Germany, they may only be used in exceptional cases, such as at the Charité hospital or military hospitals. The legal framework is strict: all phages must be GMP-certified, which makes production very complex and cost-intensive.
From this exchange, we learned that phages are already extensively researched, yet their development, production and marketing in Germany remains highly challenging and costly until this day.
The interview motivated us to further develop the economic aspects of our project Bactolyze to research alternatives to Antibiotics. Additionally we wanted to establish contact with researchers and companies such as Christine Rohde, phage researcher at the Leibniz Institute DZIF, as well as the companies Micreos and Invitris to gain deeper insights into regulatory and market-related challenges.
In our interview with Dr. Berger, we gained detailed microbiological insights, with a focus on chronic watery diarrhea inducing enteroaggregative Escherichia coli (EAEC) and uropathogenic Escherichia coli strains (UPEC) that cause urinary tract infections (UTIs) which are associated with biofilm formation. We learned about the genetic mechanisms underlying bacterial virulence and plasmid dynamics, providing valuable context for understanding the adaptability and persistence of pathogenic E. coli. The discussion deepened our comprehension of the complexity of bacterial behavior and highlighted key ethical and practical considerations relevant to the development of phage and endolysin-based therapies.
Our objective in engaging with Dr. Berger was to obtain his expert knowledge of E. coli biology such as understanding their capability of producing biofilms, their plasmid-encoded antibiotic resistance as well as of the interaction between bacteria and bacteriophages since some phages like M13 can influence bacteria conjugation. We aimed to investigate how microbial genetics and physiology influence phage therapy efficacy and draw lessons for designing realistic, clinically and relevant experimental approaches.
Dr. Berger elaborated on the genetic regulation of E. coli, pointing out that genes closer to the origin of replication tend to be more highly expressed due to copy number effects. We thought about implementing this knowledge by placing our phage DNA close to the origin of replication therefore getting a higher phage yield. Furthermore, Berger explained how fimbriae contribute to biofilm structure; K-12 E. coli produces type 1 fimbriae which mainly support moderate biofilm formation and adhesion under stress conditions while EAEC produces aggregative adherence fimbriae (AAF) that result in a stronger and greater quantity of biofilm than type 1 fimbriae. In terms of experimenting on biofilm formation and inhibition EAEC is a better suited bacteria strain than K-12 for our goal. Although EAEC is a biosafety 2 bacteria strain making it difficult for us to get access to, Berger offered us to work in his S2 laboratory at the University Medical Center of Münster.
The expertise of Dr. Berger helped us refine the focus of our project and deepened our understanding about the E. coli strains most relevant for our work, including their capacity for biofilm formation especially in K-12 and EAEC strains. Therefore, we chose to do our biofilm inhibition assays on EAEC for its capability to produce high and strong biofilm yields.
The Leibniz Institute DSMZ - Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (German Collection of Microorganisms and Cell Cultures) is a biological resource center with a focus on microbial diversity. Lately, they have also worked on various aspects of bacteriophage research, with e.g. PhageDive and Phage4Cure, projects that aim to build huge bacteriophage libraries and databases (PhageDive ) as well as advancing a defined licensed phage cocktail towards therapeutical application. Our exchange provided valuable insights into regulatory challenges, the complexity of phage therapy formulations, and current scientific challenges, such as biofilm resistance.
Our primary objective in contacting Dr. Christine Rohde and Dr. Johannes Wittmann, from the DSMZ was to deepen our understanding of ongoing initiatives regarding phage biodiversity and the challenges limiting the clinical translation of phage-based medicinal products. DSMZ was the perfect match for these matters, as they are involved in scientific research and are trying to overcome legal obstacles to bring formulated phage therapeutics to market in Germany and the EU. Additionally, we were interested in their perspective on the practical challenges of applying phages and the potential of endolysins.
DSMZ’s PhageDive initiative aims to create a comprehensive bacteriophage database by pooling a vast collection of host strains and phages as well as associated data. This effort reflects the growing importance of identifying phages with therapeutic potential. Their Phage4Cure consortial project is notable as the only currently active clinical trial on phage therapy in Germany running under the EU Clinical Trials Register, highlighting the scarcity of clinical trials in this emerging field. However, regulatory obstacles remain significant. A recent publication by DSMZ researchers points to challenges in the approval of phage mixtures, stressing that strict GMP requirements may be neither feasible nor practical in this context.
This issue becomes even more pressing in light of the limitations of antibiotics: while bacteria continually develop new resistance mechanisms, antibiotics can only attempt to keep pace. In contrast, phage cocktails or endolysins offer a fundamentally different approach — one where adaptability is on the side of the therapy rather than the pathogen. However, if regulatory barriers remain unchanged, which means that each therapy phage has to be validated through the complete traditional licensing process, the potential of these innovative treatments risks being stalled in the same cycle that has limited antibiotics.
One of the most valuable insights we gathered from the meeting was knowledge about other endolysin- and phage-related researchers and institutions that guided us on our path forward.
Building on DSMZ’s input, we identified several more potential collaborators for our project, including biofilm experts, endolysin researchers, institutions involved in clinical testing and application, and artilysin/AMP experts. We also identified biofilms as a persistent and pressing obstacle and aligned our research priorities with strategies to test phage and endolysin activity under biofilm conditions. At the same time, we acknowledged that GMP requirements pose major hurdles for phage products in the sense of active pharmaceutical ingredients (APIs), while lysins as purified proteins are comparatively easier to standardize and manufacture under GMP, making them an additional or alternative more straightforward candidate for clinical translation.
At the BFH Meetup, we presented our project Bactolyze and received valuable feedback. While our innovative work on endolysins was praised as highly impactful, we were encouraged to narrow our focus. This input led us to center our project more clearly on the endolysin production platform, supported by further expert discussions.
The BFH Meetup provided the ideal opportunity to present our project Bactolyze to the iGEM community for the first time. We received detailed and valuable feedback from four judges during our project pitch and the subsequent poster session, which we continuously implemented into our work.
All four judges evaluated our project as innovative and potentially highly impactful. Our approach to developing endolysins as an alternative to antibiotics was highlighted as a particular strength. At the same time, the judges advised us to sharpen our focus from a general endolysin platform to more specific applications to avoid pursuing an overly broad approach. Additionally, we were encouraged to further pursue our biofilm-combatting approach, as this definitely is considered to be one of the most pressing limitations in curing bacterial infections efficiently. At the same time, the concern of a possible immune response to phages was expressed. However, we received strong praise for the conceptualization of our wetlab approach and the achievement of our engineering success.
In addition, we met Dr. Patrick Grossmann , CEO and co-founder of Invitris, a startup specializing in cell-free expression systems for bacteriophages. At the meetup, we learned that they were currently starting to work on endolysin production. As our iGEM journey progressed, Dr. Grossmann became an important expert within our integrated human practices.
The overall positive feedback from both the judges and fellow iGEM teams significantly reinforced our determination to further advance and expand our project. We carefully considered the constructive criticism received and refined our project narrative by reevaluating our strategy for endolysin development. This reflection revealed a critical need for a reliable, rapid, and cost-effective platform for endolysin production, which we consequently positioned at the core of our work. Concurrently, we aimed to enhance endolysin variants by screening their lytic efficiencies and optimizing them through the application of our computational model. For prospective application methods, we also envisioned utilizing bacteriophages as delivery vehicles, onto whose surface's human-derived peptides with anti-biofilm properties would be fused to improve biofilm infiltration and disruption while at the same time lowering the chance for a high immune reaction. To substantiate this new focus scientifically, we engaged in further discussions with experts in the field.
Professor Dr. Bozzaro is a professor of ethics at the University of Münster and conducts research in applied ethics with a focus on medical and biotechnological issues. During the interview, she identified antibiotic resistance as an ethical challenge of a “limited resource” and emphasized sustainability as a moral obligation to future generations. She discussed justice-oriented approaches to responsible use, explored opportunities for alternative therapies with us such as phages, and highlighted education and awareness as crucial levers for public health.
Our objective in consulting with Ms. Bozzaro was to examine the ethical dimensions of antibiotic use and resistance from a bioethical perspective. Our objective was to ascertain how moral responsibility can be distributed between present and future generations. Furthermore, the issue of how the introduction of alternative therapies can be ethically justified and which social mechanisms from politics to medicine to individual education could foster the sustainable use of antibiotics.
Ms. Bozzaro stresses that antibiotic resistance is a societal challenge requiring cooperation across human medicine, veterinary medicine, and environmental health. She refers to the One Health concept, which underscores the interdependence of human, animal, and ecosystem health. Sustainable solutions to antimicrobial resistance demand cross-sectoral responsibility and transdisciplinary action. It is vital that political, social and scientific actors all play their part in safeguarding global health.
She frames antibiotic resistance as a genuine problem of ethical sustainability: antibiotics are a finite resource, and today’s use directly shapes the treatment options of future generations. On the topic of “grandchild generation justice,” she stresses our active responsibility toward future descendants, despite the potential of technological innovation to mitigate, but not fully resolve, the issue.
In the context of ethical sustainability, she believes that the most important strategies are to increase efficiency by making accurate diagnostics before prescription and where necessary, to ration and reduce the use of antibiotics in less urgent cases. While such measures may be challenging for society to accept, they may become inevitable in the long term.
It is essential that there is comparable scientific evidence for both standard and alternative therapies. She further emphasizes the need for political support and industrial investment to develop such alternatives and ensure equitable access.
In terms of fair resource allocation, she highlights the ethical challenges of prioritizing certain patient groups such as younger versus older patients while highlighting the role of education and health literacy. It is imperative that both physicians and patients develop a comprehensive understanding of the importance of antibiotic usage, including the necessity of responsible and prescribed dosing. Cultural differences in the prescription process introduce further complexity to the development of ethical guidelines.
We integrated strengthened education and communication as central strategies against resistance development.
Our project incorporated the One Health approach by promoting long-term sustainability through education across generations, fostering a deeper understanding of sustainability and its significance in biology.
“We have an active responsibility to our children and grandchildren. Even though innovation can solve many problems, we cannot rely on it alone, the sustainable use of antibiotics is an ethical obligation to the future.”
– Prof. Dr. Claudia Bozzaro
As part of our ethics lecture, we facilitated an interactive discussion with students on the subjects of responsibility, equitable access, and risks associated with new therapies. Furthermore this debate emphasised the collective responsibility of hospitals, society, healthcare professionals, policymakers, and scientists to gurantee the safe, transparent, and fair implementation of new therapies, as well as pointing out the critical role of education and interdisciplinary collaboration.
As part of our ethics lecture, we also wanted to gather feedback from students and discuss responsibility, equitable access, and the risks of new therapies with them, as well as get their opinions on these topics. To this end, an ethical discussion round was held towards the end of the lecture supported by interactive and anonymous Mentimeter polls.
At the outset, the question whether access to alternative therapies, such as phage therapy should be available at an early stage or reserved exclusively for seriously ill patients was discussed. Approximately one-third of those present argued in favor of restricting access to seriously ill patients, while the majority advocated for early use. During the debate, students expressed the view that cost-intensive therapies impose a burden on the solidarity-based healthcare system and therefore should be reserved for cases without alternatives. However, the argument was made that established phage therapy could be significantly more cost-effective. It was also suggested that early diagnosis and appropriate treatment would both reduce patient suffering and reduce transmission to other people or hospital staff.
With regard to the risks posed by multi-resistant pathogens versus genetically modified organisms (GMOs) used to combat them, only approximately seven percent of those present considered GMOs to be a greater problem. The majority agreed that genetic changes also occur regularly in nature. Furthermore, phages are not capable of infecting human cells and cannot be approved in Germany without strict testing procedures. As a result, they were therefore perceived as posing a lower risk and being more controllable.
In the subsequent discussion on responsibilities for the spread of multi-resistant bacteria, various actors were considered: Hospitals have a duty to consistently implement hygiene regulations, establish preventive framework conditions such as testing, and ensure infection control measures in patient care. It is the responsibility of society to comply with hygiene rules, especially hand washing and disinfection, as well as to act in solidarity. Clear information from doctors was called for to ensure implementation, and the population's own duty to inform was also emphasized. Farmers and healthcare professionals are encouraged to use antibiotics judiciously and to diversify their application whenever possible. Politicians should create suitable conditions, provide targeted incentives, and prioritize the issue of infection control. Science was viewed as responsible for making information more accessible, developing safe and efficient therapies, and communicating transparently.
Finally, we discussed the responsible use of new therapies in the future. It was emphasized that comprehensive safety studies and careful validation are necessary, as is the role of science in post-approval monitoring. At the same time, there were calls for cross-border and interdisciplinary cooperation that also takes innovative tools such as artificial intelligence into account. Politics was subject to close examination, underlining the importance of informed decision-makers who can encourage confidence in science and effectively regulate lobbying. In addition, guidelines for equitable access to new therapies for all were called for. However, the importance of education was once again emphasized most strongly.
During the discussion, the urgent need for effective guidelines for equitable access to therapies was emphasized, but it needs to work without placing an excessive burden on the solidarity-based healthcare system. For this reason, we sought the counsel of a legal professional and an insurance specialist to ascertain the current regulations pertaining to phage therapy and its potential financial underwriting. At the same time, we gained a deeper understanding of the importance of safety and sound knowledge about the mechanisms of action of new therapies for social acceptance, which encouraged us both in our further public relations work and in the specific project of organizing an art exhibition.
Invitris is a cutting-edge biotech company, originally inspired by an iGEM project, specializing in cell-free expression systems for bacteriophages. More recently, they have also begun applying their expertise to the production of endolysins. Our conversations with the team provided valuable insights into the current state of cell-free protein expression for antimicrobial research, the strong industry demand for scalable and rapid platforms, and state-of-the-art screening methodologies. The exchange also connected us with additional resources for advancing our own project, while highlighting both the technical opportunities and industrial needs shaping the development of endolysin-based applications.
Our primary objective in engaging with Dr. Patrick Grossmann (CEO and Co-Founder) and Dr. Kilian Vogele (CTO and Co-Founder) was to gain a deeper understanding of cell-free expression and its applications to phage-derived enzymes such as endolysins. Following our initial meeting with Patrick at the BHF European Meetup, we aimed to learn more about their phage expression kit, scaling potential, and assay methodologies, as well as to receive practical guidance on how their platform approach could support our own endolysin research. We were also interested in feedback on potential collaborations and in hearing about future perspectives for downstream applications and next-generation screening technologies for endolysin development.
Invitris initially developed cell-free expression systems for bacteriophages, which are already being deployed successfully and have attracted considerable industry interest. However, beyond phages, there is a clear and growing demand from multiple companies for a cell-free screening platform tailored to endolysins, reflecting the urgency of advancing endolysin research and related therapeutic development.
In terms of functional validation, Invitris emphasized that most current assays for endolysins still rely on spot assays and growth inhibition assays, as these methods provide simple, reproducible, and widely accepted ways to assess antimicrobial activity, thereby highlighting their importance for establishing robust baselines in our own experimental work.
Looking ahead, the company sees significant innovation potential in droplet-based microfluidic screening platforms, where protein expression and activity testing could be performed in miniaturized, high-throughput reactions. When combined with electrical impedance or optical density measurements, this could allow rapid, sensitive, and parallelized assessment of endolysin activity.
Finally, Patrick stressed that achieving scalability will be a critical milestone for future cell-free systems at Invitris. They aim to scale up the volume of a single expression reaction to 1–5 liters, providing a crucial bridge between laboratory-scale experiments and early industrial applications, and laying the groundwork for clinical-scale production.
Guided by Invitris’ insights, we integrated spot assays and growth inhibition assays as core evaluation methods to align our experimental design with established industry practices. At the same time, we identified droplet-based microfluidic platforms as a promising future direction for even faster and more cost efficient high-throughput screening, even though these have not yet been applied to endolysins.
This interaction not only expanded our technical perspective but also highlighted clear industrial demand for tools enabling rapid endolysin prototyping and validation. Here, Patrick also enabled us to connect with other stakeholder companies, such as Micreos, whom we consulted in further interviews.
Overall, this exchange strengthened our appreciation for how innovations in cell-free systems and next-generation screening technologies can accelerate the translation of engineered endolysins from the lab bench to therapeutic and industrial applications.
Micreos Pharmaceuticals is a pioneering biotech company specializing in engineered endolysins aimed at combating pathogenic bacteria with precision while preserving the microbiome. The discussion illuminated key challenges in clinical development, regulatory processes, and strategic focus. It also offered valuable feedback on our project idea of advancing endolysin-based therapeutics whilst especially highlighting the need for more research on endolysins against Gram-negative bacteria.
Our primary goal in reaching out to Dr. Matthew Dunne (Chief Scientific Officer at Micreos Pharmaceuticals) was to gain first-hand insights from one of the few pioneering companies dedicated to the therapeutic use of endolysins, regarding challenges to be faced on the way to a marketable product, lessons learned in endolysin engineering, as well as valuable feedback on our project idea.
Micreos Pharmaceuticals focuses on engineering endolysins for clinical applications such as the treatment of Atopic Dermatitis in dermatology with future potential in oncology, such as the treatment of Staphylococcal dysbiosis associated with Cutaneous T-cell Lymphoma (CTCL).
According to Dr. Dunne, key challenges in developing endolysin therapies include maintaining protein stability under physiological conditions, demonstrating pharmacological effects, and navigating the diverse clinical trial requirements depending on the intended application. Still, endolysins are generally viewed as very promising due to their low risk of resistance development, broad pH stability, and high efficacy against bacterial biofilms.
Currently, Micreos focuses on advancing its endolysins targeting Gram-positive bacteria. For those targeting Gram-negative bacteria, Dr. Dunne highlighted the need for advanced engineering strategies to enable penetration of the outer cell membrane, as well as improved screening methods to rapidly identify promising variants, supporting our idea of developing such a platform independent of initial production problems when using recombinant expression. Nonetheless, Dr. Dunne regards recombinant production through up-scaled and process-optimized fermentation as the currently most cost-effective strategy for clinical-scale endolysin production.
Regarding our experimental approach, he recommended conducting enzyme activity assays in relevant growth media, rather than buffer-only systems, and testing both actively growing and overnight bacterial cultures to better reflect real conditions. For protein engineering of endolysins and cationic peptides, he advised assessing natural linkers to optimize functionality.
Guided by Dr. Dunne's expert advice, we refined our project's scope to prioritize engineering of endolysins targeting Gram-negative bacteria. We also adapted our lab protocols to test growth inhibition assays in nutrient-rich media and included comparative tests using both log phase and overnight bacterial cultures.
This conversation also heightened our awareness of public acceptance considerations shaping future human practice and educational work. Overall, the interview strengthened our perspective on endolysin therapy and our practical approach towards advancing this field of research within realistic industrial and clinical frameworks.
The interview reveals that phage therapy in Germany is currently classified as an individual therapeutic trial. Phage preparations are currently not available as approved drugs. Although there are no legal differences compared to other European countries, practical implementation difficulties and a lack of compounding options hinder its use in Germany. For phage therapy to be widely adopted, further research, legal adjustments, and incorporation into medical guidelines are required. Based on this insight, we prioritized working with endolysins to promote broader application and involved medical professionals and regulatory service providers in our Integrated Human Practices to further explore this topic.
The interview with the lawyer-biologist aimed to achieve a more profound comprehension of the legal and regulatory framework governing phage therapy in Germany, and to elucidate the challenges related to approval, reimbursement, and utilization of phages and endolysins.
In Germany, phage preparations have not yet received approval as pharmaceuticals, and consequently, their utilization is not permitted as off-label use. Instead, their application falls under the remit of individual therapeutic trials. Therapeutic trials are permitted in exceptional cases and only after individual review if no alternatives exist.
The current approval hurdles and regulatory peculiarities in Germany have prevented widespread use thus far. Approval of phage therapy is often unsuccessful due to a lack of double-blind studies, high technical requirements, and comprehensive testing by the authorities. The process encompasses preclinical and clinical data, in addition to an examination of the manufacturing conditions. From an economic perspective, personalized phage therapies offer minimal commercial viability for the pharmaceutical industry, primarily due to their inability to be mass-produced. Furthermore, genetically engineered phages are subject to particularly stringent regulatory requirements, rendering their development and approval processes highly complex and financially burdensome. Consequently, the potential for financial gain in this field remains extremely limited. Conversely, endolysins have already been employed in select instances.
The legislation governing medicinal products is largely uniform across Europe, ensuring legal consistency between countries such as Belgium and Germany. However, phage therapy has been more extensively implemented in countries such as Georgia, primarily due to less stringent regulatory oversight. In Germany, extemporaneous manufacture of phages is not currently feasible.
To facilitate the widespread utilization of phage therapy, there is a necessity for legal adjustments, further studies, and inclusion in medical guidelines. Currently, there is a risk of legal repercussions for pharmacies and medical practitioners who deviate from standard therapies.
The interview revealed that the implementation of novel therapeutic approaches in Germany largely depends on the establishment of explicit regulatory frameworks and approval procedures. Based on these findings, we placed greater emphasis on working with endolysins to guarantee a more widespread application. Additionally, we decided against working with GMO phages and reconsidered using WT phages in combination with the raw peptides for biofilm tests. To further investigate this topic, we also involved medical professionals and specialized service providers from the areas of approval and reimbursement in our Integrated Human Practices.
Our interview with Vera Winkelsett from LVM Health Insurance focused on the evaluation of novel biotechnological therapies by LVM Health Insurance. The company expressed a general openness towards such innovations but emphasized the necessity of official regulatory approval and clear medical indication. In the absence of formal approval, requests are subject to individual case assessments, particularly in situations where standard treatment options have been exhausted. While the insurance company supports approaches in personalized medicine, these must be grounded in robust scientific evidence and demonstrate economic feasibility. In practice, privately insured patients may experience comparatively easier access to these therapies. However, the decisive factor for coverage remains the medical indication.
The interview partner works for the German insurance company LVM, which also offers private health insurance. The purpose of the contact was to gain insights into how innovative biotechnological therapies, such as phage therapy or therapy with endolysins, are evaluated and whether these treatments are covered for patients.
Innovative biotechnological therapies, like phage therapy or endolysins, are generally viewed positively by LVM Health Insurance; however, reimbursement is primarily linked to official regulatory approval (e.g. BfArM). Clinical guidelines from medical societies can facilitate the assessment but are not considered an absolute prerequisite. In the absence of regulatory approval, reimbursement decisions are made on a case-by-case basis, particularly when established treatment options have been exhausted. Under certain conditions, experimental treatments or therapies conducted abroad may also be reimbursed, provided that medical necessity is clearly demonstrated.
Personalized medicine is already supported in selected cases, though each request undergoes rigorous evaluation regarding scientific evidence. Access to innovative therapies tends to be more flexible for privately insured patients, yet medical necessity consistently remains the decisive criterion.
These insights illustrate that regulatory frameworks and healthcare system requirements play a pivotal role in determining whether new therapies can be integrated into clinical practice.
From the interview with LVM Health Insurance, our project derived the insight that regulatory requirements should be considered from the very beginning when developing new therapeutic approaches, since official approval is later essential for reimbursement. Building on this finding, we engaged in targeted discussions with medical professionals as well as a consultancy specializing in approval and reimbursement processes, in order to gain a clearer understanding of the relevant requirements.
The discussion with Michael, an experienced iGEMer and biotechnology student, helped us narrow down our plans for the cell-free expression of endolysin constructs and sparked the idea of molecular dynamics-based modeling of antimicrobial peptides (AMPs) for later fusion with endolysins.
The aim of this exchange was to receive conceptual guidance from Michael, a member of the Münster SynBio network (a university group consisting of iGEM Münster alumni), on the future direction of our project within the iGEM framework. Entering the discussion, we already had a strong grasp of the main challenges in therapeutic endolysin research, particularly in developing solutions against Gram-negative bacteria. We had also decided to employ the ALiCE cell-free expression system for endolysin production, recognizing several advantages over conventional recombinant expression methods. These include the potential ability to successfully express endolysins independent of their target activity, avoiding toxicity issues in bacterial hosts, and suitability for high-throughput protein screening.
During the exchange, we agreed that the foremost step must be to establish a proof of concept demonstrating that functional endolysins can be produced using the ALiCE system. We also focused on the engineering of endolysins for targeting Gram-negative bacteria with our idea to develop a modular platform approach that integrates the enzymatically active domains (EADs) of endolysins, natural linkers, and antimicrobial peptides (AMPs) for membrane penetration, enabling domain shuffling to facilitate future optimization and screening. Here, Michael emphasized the potential of using a cell-free expression system like ALiCE not merely as that but as a molecular toolbox by optimizing the system with suitable vectors tailored for high-throughput cloning methods like Golden Gate or recombinase-based systems.
Additionally, building on his suggestion, we also resolved to integrate molecular dynamics-based modeling of AMPs to gain mechanistic insights into interactions with bacterial outer membranes, drawing on the expertise of our team’s theoretical chemists. In this context, Michael also encouraged us to address potential patent restrictions not only for the AMPs but also for all biological parts utilized in our project. Consequently, we arranged a subsequent consultation with a patent attorney to clarify intellectual property considerations.
In addition to these core decisions, the discussion also considered the potential to compare different expression systems, including the eukaryotic ALiCE system, the prokaryotic cell-free expression system from INVITRIS, which is based on lysate from pathogenic E. coli strains, and recombinant expression in E. coli, particularly in terms of endolysin protein yields. Finally, Michael provided an assessment of the feasibility of scaling up production using ALiCE, an important consideration for applied contexts and larger-scale protein synthesis.
Because of our wetlab approach to couple AMPs (antimicrobial peptides) to endolysins, we considered studying the AMP insertion mechanism through MD simulations, to help our wetlab to decide which AMPs to use. Molecular dynamics (MD) simulations are computational methods that model the physical movements of atoms and molecules over time by numerically solving Newton’s equations of motion to study the structural, dynamic, and thermodynamic properties of molecular systems.
To get more insights into MD simulations, we consulted Dr. Diddo Diddens, research group leader at the Jülich Forschungszentrum, who investigates transfer and transport processes using molecular modeling.
The conversation provided insights into the basics of the MD simulation program GROMACS, technical aspects such as parallelization, and theoretical concepts, including the function and application of specific thermostats and barostats.
The objective of our conversation was to gain a deeper understanding of MD simulations and discuss some technical details of GROMACS.
MD simulations are a powerful tool for investigating mechanisms at the molecular surface. We chose the MD engine GROMACS, one of the most widely used MD programs. In our conversation we addressed the v-rescale, a modified Berendsen thermostat that generates a canonical ensemble. We also discussed the differences between “constraint” and “restraint” in GROMACS (why in umbrella sampling), as well as some other technical details. Parallelization strategies, particularly the use of OpenMP versus MPI on clusters, were also discussed. An explanation to some of these technical aspects can be found on the igem gitlab repository and on the modeling page.
Following our conversation, Dr. Diddens introduced me to two PhD students in Prof. Heuer’s group at the University of Münster, Annemarie Quas and Clara Rickhoff, who are investigating membrane properties with MD simulations.
The insights concerning MD simulations with GROMACS we obtained from this conversation were instrumental in establishing our MD simulations to study the insertion of AMP into the outer membrane of Gram-negative bacteria. In addition, the conversations we had with Annemarie Quas and Clara Rickhoff led to a fundamental analysis tool being used for our model.
In Germany, physicians are bound by strict guidelines when treating antibiotic resistant infections. Alternative therapies, including phage therapy or endolysins, can only be prescribed following regulatory approval and recognition by health insurance providers – a process that requires regulatory change, partly driven by social pressure. At the same time, biofilm-associated implant infections represent a serious challenge, highlighting the urgent need for alternative therapeutic solutions.
With this interview, we sought to understand how antibiotic resistance is addressed in medical education, what preventive measures are implemented in Germany, and the extent to which physicians are free to prescribe alternative or experimental therapies. Therefore, we interviewed two medical students from Munich, one in her final semester and one shortly before his final state examination.
Antibiotic resistance is a central topic in medical education, with a strong emphasis on ‘antibiotic stewardship’ to ensure responsible use. However, phage therapy was not taught as an alternative. Physicians are bound by treatment guidelines: if an antibiotic fails, an antibiogram determines the next drug to be prescribed. While the students had not yet encountered cases where no reserve antibiotic was effective, multi-resistant cases are increasing in Germany, and some infections may soon have no effective antibiotic.
Deviating from guidelines carries legal risks, as physicians can be sued by patients or health insurers, so therapies outside standard protocols are rarely used. For phage therapy to become a viable treatment option, official regulatory approval, inclusion in clinical guidelines, reimbursement by health insurers, and economic justification are required. Both students believed regulations could change under sufficient pressure, for example in the event of a large-scale health crisis.
They also highlighted biofilm-associated implant infections as a prevalent and critical clinical challenge in Germany. During their studies, they encountered cases of implant-associated inflammation weekly, often requiring surgical reopening and rinsing of wounds. Once surrounding tissue is affected, treatment options are extremely limited, sometimes resulting in amputations.
We learned that medical treatment in Germany operates under stringent regulatory frameworks. For phage or endolysin therapies to become realistic options, regulatory approval, reimbursement by health insurance providers, cost-effectiveness, and broader social acceptance are essential. This insight motivated us to expand our public engagement and explore approval pathways for alternative therapies. It also strengthened our focus on antimicrobial peptides (AMPs) against biofilms, which show particular promise in applications such as rinsing solutions.
“Man-made problems (and restrictions) can also be changed by humans if the incentive is great enough.”
– Medicine students
Annemarie Quas and Clara Rickhoff are two PhD students who are working on membrane simulations in Professor Heuer’s group at the University of Münster. We had two short conversations over the time span of the whole iGEM project. In our first conversation, we discussed why the Potential of Mean Force (PMF) calculated from our umbrella sampling did not correspond to the results in the work of (Sharma and Ayappa, 2022). PMF essentially represents the free energy profile along the chosen reaction coordinate and therefore crucial for insight into the relative stability of states and energy barriers in the system. The second conversation was about our unbiased simulations and which membrane properties we can analyze based on these.
After our conversation with Dr. Diddo Diddens, he introduced us to two PhD students who are working on membrane simulations in Professor Heuer’s group at the University of Münster.
The objective of the first interview was to understand why the umbrella samplings couldn’t reproduce the results in the paper (Sharma and Ayappa, 2022). The second was to find out which properties could determine membrane instability induced by the AMPs.
In our first conversation, they suggested studying the densities of the system over a simulation time of 1 µs to compare the densities with the paper of Sharma and Ayappa (2022). This led us to continue the unbiased simulations after 1 µs, to see if the peptide inserts even deeper than in the O-antigen (OA) region of the membrane.
During our second conversation, Annemarie recommended that we use the Python package “MDAnalysis” to analyze peptide properties during the simulation. Furthermore, she recommended reading the work of Triparthy et al. (2020) on 3D lipid packing defects, which could be an interesting membrane property to study (Tripathy, Thangamani and Srivastava, 2020). A more detailed description of the lipid packing defects can be found on the modeling page.
The first conversation led us to perform an unbiased simulation with a simulation time of 4 µs, after observing an insertion into the OA region of the membrane at 1 µs. The second conversation led us to study lipid packing defects with “PackMemb”, which analyzes 2D instead of the more complex 3D lipid packing defects, because we still needed to adapt the package to our Lipid A rather than the phospholipids usually analyzed by this package (Gautier et al., 2018). Therefore, we did not have time to study the more realistic 3D lipid packing defects. In hindsight, analyzing the 2D lipid packing defects was already enough to observe a significant difference after the insertion of the peptide into the sugar region of the membrane.
In a brief legal consultation regarding HY-133, the first engineered endolysin to enter clinical trials, we sought clarification on the legal boundaries of using HY-133 in our research framework for proof-of-concept in vitro production of functional endolysin. The discussion confirmed that, while academic non-commercial research might generally be possible, the situation is legally complex and cannot be guaranteed to fall under a research exemption. Including HY-133 in our project would therefore carry potential risks with limited additional benefit. Based on this assessment, we decided not to pursue HY-133 for our project.
We consulted a patent attorney to determine whether HY-133 or any other patent-protected components could be employed in our laboratory for research purposes and whether submitting an adapted version of the HY-133 gene sequence as a part for the iGEM registry would be legally feasible.
In Germany, there is no general statutory exemption that allows unrestricted use of patented materials for research purposes. Although § 11 (2) PatG (German patent law) stipulates that “The effect of the patent does not extend to acts for experimental purposes relating to the subject matter of the patented invention”, this exception requires careful case-by-case interpretation and was not comprehensively reviewed for our project. HY-133 falls under a European patent covering both the enzyme sequence and specified pharmaceutical formulations (e.g., HEPES buffer and excipients). According to legal advice, there remains a tangible risk that even good-faith academic applications could inadvertently fall within the scope of the granted patent.
Given this, direct submission of HY-133 to the iGEM parts registry was deemed legally possible but strategically unnecessary, offering limited benefit while carrying potential risk.
Based on this guidance, we decided not to include HY-133 in our project design or part submissions. Instead, we focused on endolysins that are free of patent restrictions. This experience improved our understanding of the intersection between patent law and synthetic biology, enabling us to more accurately assess the risks and benefits of working with patented biological materials.
Dr. Hester explained that the process of transporting water-soluble proteins, such as endolysins, using nanoparticles is complex, as these proteins must be protected from stomach acid. Other targeted drug delivery would require expensive, long-term research, so she recommended phages as a faster and more cost-effective alternative. Potential risks, such as hemolytic effects, were discussed and reinforced our decision to focus on phages rather than nanoparticles.
We contacted Dr. Hester, group leader at the Institute of Pharmaceutical Technology and Biopharmacy at the University of Münster, to learn more about alternative transport methods for endolysins and antimicrobial peptides (AMPs). Her research focuses on improving the bioavailability of molecules in the body using nanoparticles.
Dr. Hester outlined the broad range of nanoparticle systems available but stressed the challenges associated with transporting water-soluble proteins, such as endolysins and AMPs, using nanoparticle technology. For intestinal application, pH-resistant formulations would be needed to protect the proteins from stomach acid.
To achieve a sufficient concentration of active substance at the bacteria, the nanoparticles would need ligands that bind specifically to bacterial surface molecules. She also recommended a targeted release system that only releases active ingredients once the nanoparticles attach to bacteria. However, developing such a system is costly and can take several years. Therefore, she recommended sticking to phages for faster utilization.
Pills offer a simpler alternative but only allow pH-controlled release in the intestine, which lacks precision, and require a high concentration of the active ingredient. Dr. Hester also highlighted potential toxic or hemolytic effects of peptides if nanoparticles cross the intestinal barrier. For possible pulmonary application, microparticles are preferred as they are not easily exhaled. She noted that administration via the nose should be avoided due to the risk of active ingredients entering the brain.
The interview demonstrated the significant complexity of nanoparticle technology. Given the short research time and our objectives, we decided to focus on phages as transport systems to reduce production costs. We also considered the risk of hemolytic effects, which led us to discontinue the use of the SMAP29 peptide. Overall, the interview reinforced our focus on phages over nanoparticles.
“If you already have a good working system (phages), don’t make it more complicated than needed.”
– Dr. Sarah Hester
Our discussion with Josua Janowski from Diapharm provided us with a comprehensive and well-founded evaluation of the current regulatory landscape for phage therapy. Diapharm emphasized the current challenges, but also highlighted the increasing regulatory momentum, driven by newly published EMA (European Medicines Agency) guidelines and additions regarding phage therapy to the European Pharmacopoeia. Overall, the company expressed a positive outlook on the emergence of a viable framework for approving phage therapeutics in Germany and Europe within the medium term.
We consulted Diapharm to identify the regulatory requirements and potential obstacles relevant to the approval of our phages, endolysins, and antimicrobial peptides (AMPs). The objective was to proactively identify possible challenges and obtain a realistic appraisal of the feasibility and procedural aspects from an experienced regulatory consultancy.
Diapharm, a consultancy specializing in regulatory support in the healthcare sector, guides pharmaceutical manufacturers throughout the entire approval process, from dossier preparation to market authorization. During the interview, they provided current perspectives on phage preparations and endolysins, acknowledged as innovative alternatives to antibiotics, yet encountering significant regulatory barriers.
To date, phage-based medicinal products have not been authorized across Europe, with applications predominantly limited to compassionate-use or exceptional cases. In Germany, phage therapy is currently authorized solely as an ultima ratio treatment, meaning it is only considered when all other therapeutic options have failed. Additionally, the approval process for phage therapeutics is notably more demanding than for conventional antibiotics or inactivated vaccines, and major regulatory authorities including the Federal Institute for Drugs and Medical Devices (BfArM), the Paul-Ehrlich-Institute (PEI), and the European Medicines Agency (EMA), play key roles in the review process.
Encouragingly, recent developments signal increased regulatory attention with the publication of specific EMA guidelines and the addition of a dedicated chapter on phage therapy to the European Pharmacopoeia. Despite the ongoing absence of a comprehensive legal framework in Germany, Diapharm expressed optimism, referencing advances in neighboring countries like Belgium, where phage therapy is more established. Should the German regulatory environment remain restrictive, commercializing products in other European countries with higher demand remains a viable strategy.
Encouraged by Diapharm’s positive outlook on future regulatory pathways, we have continued to advance our project involving bacteriophages. To mitigate potential approval challenges, we also intend to evaluate non-genetically modified phages in combination with pure antimicrobial peptides (AMPs), leveraging their synergistic potential while navigating evolving regulatory landscapes.
In the interview with Prof. Christine Heilmann from the Institute of Medical Microbiology at the UKM Münster, we gained insights into the comparative therapeutic potential of endolysins versus autolysins, as well as challenges and strategies in enhancing endolysin efficacy through protein engineering. The discussion further highlighted the importance of robust delivery mechanisms in achieving clinical effectiveness of endolysin-based therapies.
Our goal was to understand the therapeutic advantages and limitations of endolysins and autolysins (the proposed evolutionary precursor of endolysins), the protein engineering considerations for creating effective fusion constructs with antimicrobial peptides, and the clinical translation challenges associated with the targeted delivery of endolysins.
Prof. Heilmann confirmed our literature-based understanding that both endolysins and autolysins share a modular structure with bacteriolytic activity, being produced intracellularly before targeting the bacterial cell wall. While autolysins are subject to strict regulation to prevent uncontrolled lysis of the producing population, their activity can also be detected when applied extracellularly, although systematic studies on this remain limited. Endolysins, in contrast, have further evolved in the phage context and are generally regarded as more promising, or at least more widely explored, for therapeutic application, although the precise differences in their effectiveness remain an open subject of research.
Further, engineering endolysins fused to antimicrobial peptides (AMPs) was discussed as a promising route to enhance antibacterial activity, especially against Gram-negative bacteria. In this regard, Heilmann emphasized the necessity for careful design of linker sequences and overall protein architecture to preserve the individual and combined activities. Both fusion proteins as well as their separate components (endolysin and AMP alone) should be evaluated to identify optimal combinations.
Lastly, Heilmann proposed that, for clinical applications, developing robust and targeted delivery systems will be essential to ensure that endolysins reach bacterial infection sites in effective concentrations and maintain stability, addressing a key hurdle in translating in vitro potency into therapeutic efficacy.
This interaction informed our design strategy to focus on protein engineering that considers optimal linker design for AMP-endolysin fusions and emphasized testing both fused and separate antimicrobial components. We are also prioritizing exploration of advanced delivery platforms to improve clinical applicability of our endolysin candidates.
In an in-depth interview, Prof. Dr. Ulrich Dobrindt emphasized the unique relevance of bacteriophages as a research focus and as promising tools for modern antimicrobial strategies. He emphasized that bacteriophages, which have long been studied for their roles in horizontal gene transfer and mobile DNA elements, are once again emerging as key players in combating pathogenic bacteria and addressing antibiotic resistance at the population level. The discussion touched on the complex dynamics between bacteriophages, conjugative plasmids and bacterial resistance.
Our primary goal in speaking with Prof. Dr. Dobrindt was to gain a microbiological perspective on the practical opportunities and limitations of phage-based interventions. We sought his insights on how phages can disrupt horizontal gene transfer and their potential for real-world application in One Health settings. We also wanted to understand the critical role of education and cross-sector awareness.
Prof. Dobrindt emphasized that bacteriophages are not only regarded as new and exciting tools in modern microbiology, but also play a key role in limiting the horizontal spread of resistance genes by interfering with plasmid conjugation. He referenced their long research history, noting that as early as the 1970s, scientists around the world were studying phages such as M13 for their ability to bind bacterial F pili and block plasmid transmission, thus demonstrating the relevance of phages to areas far beyond traditional pathogen research.
Dobrindt described the One Health perspective as essential, which is an approach that recognizes the health of people, animals, and ecosystems are closely connected and interdependent. Stating that any sustainable reduction in antibiotic resistance requires the involvement of the entire population, including the agricultural, human medical and environmental sectors. Humans can acquire resistant bacteria through the food chain, for example from chicken, so efforts to reduce selection pressure and improve collective stewardship are crucial.
According to Prof. Dobrindt, phage therapy offers significant safety benefits as phages do not infect humans, and can be highly effective in reducing Campylobacter infections in poultry, for example, or through authorized applications in countries such as Poland and Belgium. However, he added that a combination of phages and antibiotics often yields the best results, particularly against biofilms.
Additionally, he cautioned that research must continue to address unknown factors, such as the potential side effects of inhibiting gene transfer and the impact of phages in environmental settings. Public acceptance of phage therapy depends greatly on how it is framed (it is preferable to use the term 'bacteriophages' rather than 'bacterial viruses'), and intensive outreach is needed. He referenced surveys showing high acceptance among academics, but less awareness in the general population.
Taking Prof. Dobrindt’s input into account, we strengthened the One Health integration in our project by actively involving partners from across all fields like industry, academia, and the general public. We refined our outreach strategy to specifically address public perception of phages. Furthermore, we were further confirmed that M13 phages are the right choice for our project, due to its potential in inhibiting the conjugation of bacteria, thus limiting horizontal gene transfer of resistance genes.
“Phages are exciting — they’re making a comeback for good reason. But to truly leverage their potential, we must educate people, collaborate across sectors and use phages where they are effective, always considering the wider system beyond human medicine.”
– Prof. Dr. Ulrich Dobrindt
During our multiple exchanges with our advisor Dr. Sriram Kumar, an iGEM safety committee member and postdoc at the University of Münster, we received valuable guidance on both the scientific framework and the safety integration of our project. His feedback helped us refine our storyline, strengthen the connection between different wet-lab work packages, and clarify how our project contributes to addressing the global challenge of antimicrobial resistance. He also provided valuable insights on human practice outreach, project framing, and biosafety in regard to working with BSL-2 strains in the iGEM context.
We reached out to Sriram to obtain feedback on three primary aspects:
Regarding general project safety, we discussed various aspects of the project, including the initial laboratory work involving phages, endolysins and AMPs, with respect to exposure and potential release, as well as future applications in nature and, primarily, in or on the human body. Regarding biosafety and responsibility when working with BSL-2 strains, Sriram emphasized the importance of carefully considering disinfection and waste disposal methods, and of adhering to local biosafety regulations. He also insisted on the need of referencing authoritative sources on german biological safety regulations in regard to biological safety, to ensure the thoroughness and accuracy of the risk assessment process. Additionally, he helped us find measures, such as adapting working volumes, for mitigating risks regarding our work with AMPs exhibiting haemolytic activity.
On narrative development, Sriram emphasized storytelling as a decisive element for impact. He recommended framing our project by moving from the broad challenge of antimicrobial resistance (supported by WHO data and local case studies) towards our specific innovation. This layered approach combines global urgency with local relevance, creating a compelling storyline for both scientific and public audiences.
From a scientific integration standpoint, he encouraged us to foreground our endolysin fusion concept and link it directly with modeling results rather than focusing solely on its implementation in cell-free systems, to strengthen the connection between theoretical design and experimental validation.
Regarding methodology, Sriram suggested beginning with simple proof-of-concept assays at single concentrations, with progression to checkerboard or gradient assays to gain deeper, clinically relevant insights.
Finally, he was among the first to encourage us to enhance our educational outreach, emphasizing the importance of communicating the project’s scope and impact in a clear and engaging way to spark interest and ensure that target audiences can readily grasp the project's broader relevance.
Sriram's feedback directly shaped key aspects of our project implementation. His guidance on antimicrobial peptides and BSL-2 strains enabled us to integrate the required safety measures ensuring approval of our iGEM Check-In forms and allowing us to proceed as planned. Building on the confidence gained from this exchange, we also invited him to deliver a biosafety talk at our iGEM Meetup, extending access to his expertise to the broader community.
Beyond safety, his advice also influenced how we communicated our project. By incorporating his recommendations on narrative structure, we linked the global challenge of antimicrobial resistance to our specific therapeutic innovation, leading to a more coherent and compelling project. This strengthened both the scientific credibility of our work and its translational relevance.
In our interview with Prof. Dr. Vincent Fischetti, a leading pioneer in endolysin research, we gained valuable first-hand insights into the current landscape and future prospects of endolysins as potent therapeutic agents against multi-resistant bacteria. We also received constructive feedback and reinforcement of our project approach.
Our goal in reaching out to Prof. Dr. Fischetti was to deepen our literature-based understanding of the therapeutic advantages of endolysins compared to phage therapy and antibiotics, the main scientific and translational challenges for clinical application of endolysins, as well as the design principles for optimizing lysins, particularly for Gram-negative bacteria. We also aimed to learn about strategies for overcoming concerns about the immune response and for improving the general public acceptance towards endolysins and phages. Lastly, we wanted to know more about potential applications beyond human medicine.
Overall, Prof. Dr. Fischetti emphasized the robustness and emerging clinical readiness of Gram-positive lysins. However, he also highlighted the significant challenges presented by the outer membrane barriers of Gram-negative bacteria. He presented promising engineering strategies to overcome these challenges, including fusion of lysins with cationic peptides. Here, the discussion underscored the essential role of bioinformatic tools and high-throughput screening in lysin discovery, optimization, and mechanistic understanding.
Prof. Dr. Fischetti also emphasized that no bacterial resistance to lysins has been observed to date, despite extensive testing, due to them targeting essential cell wall structures. This supports their strong potential not only for therapeutic use but also for prophylactic applications in infection prevention.
In the end, Prof. Dr. Fischetti pointed out that increased funding and greater visibility are essential to overcoming scepticism and awareness gaps in both the scientific community and the public, especially given endolysins’ superiority as simple, highly specific enzymes that can target dormant bacteria and better preserve native microbiota compared to whole phages.
Prof. Dr. Fischetti’s feedback reinforced the relevance of our project idea and highlighted the strength of our modeling approach, which aligns with promising strategies for engineering lysins against Gram-negative bacteria. His perspective also encouraged us to place greater emphasis on the educational aspect of our project, to help raise awareness and foster acceptance of lysins within both the scientific community and the public.
“Fusion of cationic peptides [to endolysins] is one of the most promising engineering approaches for targeting Gram-negative bacteria, even though more research is needed to fully understand exact molecular interactions […] making your modeling approach really exciting.”
– Prof. Dr. Vincent Fischetti
Our Human Practices approach ensured that scientific innovation was never viewed in isolation. Through a detailed stakeholder analysis, we identified, evaluated, and purposefully included a wide network of relevant actors. The continuous feedback from these stakeholders guided all major decisions during the project. Thus, our initial idea of using bacteriophages as an alternative to antibiotics evolved into a modular platform for the identification and optimization of endolysins and antimicrobial peptides (AMPs). Additionally, we explore combining AMPs with natural bacteriophages as a potential approach for endolysin delivery, especially in the context of biofilms, to combat resistant pathogens.
Integrating diverse perspectives was essential to our project’s development. Discussions with medical professionals emphasized the urgent need for new therapeutic approaches and reinforced the importance of advancing research in this field. It also became clear that cost-effective and user-friendly alternatives are crucial to enable real-world application in various contexts. Throughout the project, we therefore explored different application possibilities to make our technology adaptable and practical for different use cases.
Conversations with healthcare providers drew attention to the challenge of biofilms, particularly in relation to implants—an insight that significantly inspired our AMP–phage combination strategy. Discussions with industry partners such as Micreos and Invitris highlighted the importance of focusing on Gram-negative bacteria and underlined the role of public acceptance in such novel therapies.
Dialogue with experts in regulation and ethics helped us understand regulatory requirements and integrate them early into our project design. Consequently, we refined our concept—moving from genetically modified phages to combinations of natural phages, AMPs, and anti-biofilm peptides. Our interview with Dr. Claudia Bozarro emphasized the importance of education in the context of developing and implementing new strategies to combat antibiotic resistance.
Scientific collaborations also greatly advanced our project. Prof. Berger provided key insights into pathogen-related biofilms, leading us to conduct experiments in an S2 laboratory. Prof. Dobrindt emphasized the relevance of preventing bacterial conjugation, while Michael Jöst, Dr. Diddo Diddens, and others contributed detailed feedback on the MD-based modeling of AMP–endolysin fusion proteins. Additionally, Micreos and Invitris offered valuable input for designing and refining our growth inhibition and plate spot assays.
All interviews, analyses, and project adaptations were transparently documented, providing future iGEM teams with a solid foundation for their own Human Practices work. This open documentation supports method transfer and promotes continuity between future projects.
Our project demonstrates that innovative biotechnological solutions can only succeed through interdisciplinary collaboration and openness to continuous feedback. We learned that integrating perspectives from science, industry, ethics, and society is key to developing a forward-looking, responsible, and applicable project.
In the next steps, our expression platform should be further developed and expanded to other cell-free expression systems to enable the reliable production of functional endolysins and fusion proteins. Additionally, synergistic effects of AMPs, Endolysins, Phages and Antibiotics should be further investigated.
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