Human Practices
If a team is embarking on an itch-relieving project in order to produce an active ingredient that cream companies will be able to use in the future to maximize the effectiveness of their product, thereby helping patients suffering from pruritus, then they need to involve all the actors involved in the process to ensure that the future product can best serve the interests of all parties. To do this, we enlisted the help of the Faculty of Pharmaceutical Engineering at the University of Szeged on the research side, worked with a local cosmetics company and research institute on the manufacturing side, and then teamed up with other iGEM teams to produce a joint handbook.
Short Overview
Scientists and Drug Developers
We worked closely with experts in fields like pharmacology, synthetic biology, and drug development. By consulting these professionals, we could make sure our model is scientifically valid and practical for everyday use. Their knowledge of molecular processes and the specialized experimental methods were crucial to maximizing the effectiveness of our antihistamine ointment.
Drug/Supplement Producers
To make a laboratory material become a complete product, we collaborated with pharmaceutical and supplement manufacturers. These industry professionals helped us to scale up production, met requirements and ensured that the final product was safe and high-quality. Their expertise will be essential to overcome the challenges of bringing our product to market.
Doctors and Patients
A significant part of our Human Practices involves collecting feedback from healthcare providers, including dermatologists and general practitioners, as well as from potential users. Their knowledge of conditions that cause itching, like eczema allergies, pancreatitis, diabetes, or fatty liver disease helped us in developing our ointment to meet actual patient needs.
Contact with other IGEM Teams
Scientists and Drug Developers
In the process of designing our genes we consulted with Zsuzsanna Györfy, PhD, senior research associate of HUN-REN Biological Research Centre, Szeged about codon optimisation.
We have learnt the following:
- There are 6-7 rare codons in e.coli (significant above ~8%)
- rare codons play a role in the regulation of protein-folding → codon optimization is not ideal in all situations
- Recommended use of lac operons
- Recommended use of Sixpack e.coli → as experimental control
- Recommended use of pETDUET expression vector
- Export signal not recommended
- Questions/doubts:
- Measurement of enzyme activity
- Why is this better than DAO tablets?
- There are better, stronger anti-inflammatory agents.
After the consultation, we have decided to use the pETDUET plasmid, and we have appropiated our (codon optimised) genes to be able to be ligated properly into the vector, and we have decided not the use the pelB tag team HKUST 2022 had used, for technical reasons.
We are incredibly grateful for Zsuzsanna Györfy’s expertise and for helping us navigate the technical challenges of our project.
We collaborated with the Faculty of Pharmacy at the University of Szeged to help package our enzymes and assist with their transfer process. At the faculty, we tested different liposome formulations to find the best option for our project, into which we then packaged our active ingredients. Afterward, we conducted experiments to see how much of these ingredients could pass through the skin. To measure this, we used both ex vivo methods and a skin PAMPA experiment. Additionally, the Faculty of Pharmacy provided access to their plate reader, which we used to measure our aptamers.
The suggestions made by Dr. Gábor Katona and Dr. Szilvia Berkó that made our experiments more effective:
- The composition of the liposomes used in our experiments
- Using hydrogel instead of other creams, making our liposomes stable for a longer period
- Performing ex vivo experiment to get more properly validated results
- Carrying out skin PAMPA experiment to collect more data in connection with skin penetration
We would like to extend our deepest gratitude to the Faculty of Pharmacy at the University of Szeged for their invaluable support and collaboration throughout the course of our project.
The following people from the faculty were involved in our work:
Prof. Dr. Ildikó Csóka
Dr. Anita Kovács
Dr. Szilvia Berkó
Dr. Mária Budai-Szűcs
Dr. Gábor Katona
Prof. Dr. István Szatmári
Dr. Dóra Hegedűs
Prof. Dr. István Zupkó
Szalai Andrea
Dr. Zsuzsanna Schelz
Drug/Supplement Producers
In our recent meeting with Edina Pallagi, she provided an overview of her cosmetic laboratory, where she showed the sustainable practices she has implemented to minimise environmental impact. With her enormous expertise in pharmaceutical technology and science, Edina gave us valuable insights into the formulation of the ingredients used in her products, explaining how each component contributes for the final result. Additionally, she highlighted the importance of selecting responsible packaging solutions that align with sustainability while ensuring product integrity. Throughout the discussion, Edina highlighted the importance of following legal regulations and industry standards, making sure her products meet top safety and quality requirements while also complying with all the rules in the cosmetic and pharmaceutical industries.
| Name | Recommendation | How did we apply it? |
|---|---|---|
| Edina Pallagi PhD | She highlighted how much longer the approval process is for a medicinal active ingredient compared to when it is intended as a cosmetic active ingredient | We selected the raw materials taking into account the need to comply with the cosmetic authorisation rather than the medical product authorisation |
| Edina Pallagi PhD | It is difficult to change the sourcing of raw materials, and it requires a lot of work to select the supplier appropriately | We have carefully considered all options for the supply of raw materials to select the best one in every respect |
| Edina Pallagi PhD | It is unnecessary to add essential oil to the active ingredient | The planned essential oils were discarded because they were unnecessary and could have endangered stability and limited usage |
| Edina Pallagi PhD | Investors mainly approach us, as active ingredient developers, to develop the material they require | In our business plan, we focused on having good research and development to attract investors |
| Edina Pallagi PhD | According to her experience, the Hungarian market is very small, so it is worth seeking a larger market. | As a result, we examined the European and world markets as potential markets and found that we can operate most economically in the world market |
| Edina Pallagi PhD | When it comes to enzymes produced by bacteria, it is very important to pay attention to proper purification to ensure that no potentially harmful substances remain in them for the human body | In selecting the bacterium, we paid attention to choosing one that is not harmful to the human body and has already been used for enzyme production |
In a recent meeting with István Kiss, the director of BAY-BIO’s biotechnology department, he provided us with valuable insights into biotechnology. We had several questions about DNA synthesizers and fermentors, essential for the industrial production of our active ingredient, which he answered with great patience and expertise. With his knowledge in biotechnology, István offered detailed information for our production planning, uncovering areas we hadn’t considered before. Later in the meeting, he invited us for a tour in the research institute where he works. That same week, we visited the Bay Zoltán Research Institute in Szeged, where we saw a fermentor in person and had the opportunity to ask more specific questions about its operation, costs, and maintenance. The answers we received were incredibly valuable, allowing us to incorporate this new knowledge into our business plan and make it more accurate.
| Name | Recommendation | How did we apply it? |
|---|---|---|
| Dr. István Kiss | He raised the question of whether it is worth buying the machines or if renting them would be more cost-effective | After careful research, we decided it's better to purchase the machines with a large initial investment rather than renting them. |
| Dr. István Kiss | He highlighted how important the role of storage is. | We examined under what conditions the active ingredients remain the most stable, and we found that they require refrigeration. Therefore, we also designed a refrigerator system for the inventory area in the warehouse. |
| Dr. István Kiss | The fewer people there are, the lower the cost. | At first, we wanted to hire a lot of people right from the beginning, but after the meeting, we reduced the number of people to the point where they can still work efficiently without being unnecessarily too many. |
Doctors and Patients
Chronic pruritus, or persistent itching, is a significant issue faced by patients with various conditions, including chronic kidney disease, diabetes, and dermatological diseases. Accurate measurement of pruritus severity is crucial for understanding its impact on a patient's quality of life and for developing effective treatment strategies. In the context of the iGEM competition, our project focuses on addressing pruritus through innovative biological approaches and tools.
Our team has developed a series of comprehensive pruritus assessment questionnaires based on existing validated tools. These questionnaires are tailored to assess itching across different patient populations, including those undergoing dialysis, and individuals with pancreatitis, diabetes, or fatty liver disease. The tools included provide a multidimensional view of pruritus, evaluating not only itch intensity but also its impact on daily activities, sleep, and emotional well-being.
The theoretical background to our questionnaire:
The same self-evaluation quastionnaire available online: