Aim of Contact

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.

Key insights

The discussion yielded several important insights that guided the evolution of our project:

• The panel recommended considering therapeutic strategies that combine different endolysins to prevent the emergence of resistance
• Rather than beginning with a de novo endolysin, the panel advised starting with a well-characterized, known endolysin to reduce uncertainties.
• Attention should be paid to the potential immune response against phages, which could limit their effectiveness.
• The use of phages in cancer therapy was flagged as problematic due to biological complexity and ethical concerns.
• The production of phages could be associated with high costs, raising questions of scalability and feasibility.

Implementation of feedback

The expert input significantly influenced the development of our project. Based on their recommendations, we made several adjustments:

• We revised our initial plan and decided to begin with a known endolysin rather than designing a new one from scratch.
• Considering the concerns raised, we decided not to pursue the application of phages in cancer treatment.
• We further assessed the potential challenges surrounding phage production and immune response and integrated them into our ongoing risk analysis and design considerations.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of contact

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.

Insights

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.”

Implementation

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

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of Contact

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.

Insights

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.

Implementation

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.

Aim of Contact

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.

Insights

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.

Implementation

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.

Aim of Contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of Contact

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.

Insights & Implementation

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.

Aim of contact

The objective of our conversation was to gain a deeper understanding of MD simulations and discuss some technical details of GROMACS.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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.

Aim of contact

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.

Insights

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.

Implementation

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

Aim of Contact

We reached out to Sriram to obtain feedback on three primary aspects:

• how to integrate safety, biosafety, and security perspectives into our project design and documentation especially when working with BSL-2 strains and potentially haemolytic antimicrobial peptides (AMPs),
• how to effectively communicate the urgency of multidrug resistance through a compelling narrative for both scientists and the public,
• and how to enhance our human practices work by identifying target groups as well as structuring our education efforts.

Insights

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.

Implementation

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.

Aim of Contact

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.

Insights

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.

Implementation

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