Executive summary

Existing methods for microplastic detection often rely on visual classification and additional instruments, which are not suitable for nanoplastic detection, leading to potential miscalculations. Combining various techniques has shown promise but has also introduced comparability issues due to methodological differences. LuciPep combines molecular biology, bioluminescence and smartphone technology to address the challenges of microplastic detection. Its usefulness could spread beyond environmental research and monitoring and into the field of human health. Our innovative molecular method for direct microplastic quantification in raw, unpurified samples has the potential to extend to nanoplastic detection. Thus, LuciPep offers solutions to current shortcomings in microplastic diagnostics while adding significant value to the market.

LuciPep simplifies field sampling, minimizes sample treatment and provides more reliable estimates of particle concentration. Additionally, LuciPep aims to make this technology accessible to smaller laboratories worldwide, including those with limited resources, by utilizing smartphones for detection. This opens up various use-case scenarios, from environmental research institutes to water resource management agencies. Collaborations with research institutions, environmental organizations, biotech companies, government agencies and analytical laboratories will play a crucial role in refining and validating LuciPep's technology, ultimately contributing to the global understanding of microplastic pollution and its management.

LuciPep leverages molecular elements, including microplastic binding peptides (MBPs) for high-specificity detection and NanoLuc luciferase for bioluminescent biosensing coupled with smartphone-based data analysis. MBPs unique sensitivity to nanoplastics, smaller sample requirements and adaptability set LuciPep apart. It not only fills a critical gap in microplastic monitoring but also opens doors to applications in health research, specifically microplastic detection in blood samples. LuciPep's potential strengths include user-friendliness, adaptability and inclusivity, while external opportunities lie in the growing global awareness of microplastic pollution. Potential collaborations with governmental and environmental agencies, research institutions and water management authorities will further its reach and credibility. However, challenges include ensuring accuracy, staying ahead of competitors and adapting to evolving biotechnology. Effective risk management through quality control, innovation, adaptability and regulatory diligence will be essential for LuciPep's success in both environmental and health research applications.

Microplastics necessitate innovative detection methods and LuciPep emerges as a potential game-changer. Traditional techniques like microscopy lack sensitivity and spectroscopy, while effective, is costly and expertise-dependent. LuciPep's smartphone-based system offers user-friendly, cost-effective and accurate microplastic detection, making it highly accessible for both rural and urban areas. Its implementation could revolutionize waste management, enabling real-time monitoring, crowdsourced databases and substantial cost reductions by eliminating the need for specialized equipment. Moreover, LuciPep's adaptability extends its utility to studying microplastic bioaccumulation in humans, addressing health concerns. Despite potential limitations related to plastic compatibility, calibration, data handling and standardization, LuciPep's accessibility and versatility position it as a promising solution for mitigating plastic pollution and understanding its impact on ecosystems and human health.

Long-term impact

Microplastics are prevalent pollutants affecting ecosystems. Plastics break down into micro- and nano-sized particles through natural degradation processes like UV radiation or commercial processes like thermal degradation. Plastics are some of the most ubiquitous materials in human applications because of their versatility, cost-effectiveness, durability, biocompatibility, customization flexibility, chemical resistance and ease of production. Nevertheless, their disproportionate use has also made them ubiquitous in both aquatic and terrestrial ecosystems (Yee et al., 2021).

The LuciPep detection system could be a revolutionary technology in the field of microplastic detection. Conventional detection techniques involving the use of microscopy lack the necessary accuracy and sensitivity to detect minute plastic particles. Spectroscopy, although convenient for testing smaller microplastic particles, is a generally expensive alternative that also requires expertise. This hinders its use as a routine solution. LuciPep’s ease of detection and simplified smartphone application set it apart from conventional methods. Smartphones have advanced cameras and sensors that could detect sensitive signals with good accuracy. Moreover, these devices are continuously improving, which is all the more promising for our product.

The implementation of LuciPep in both rural and urbanized landscape could have a significant impact on the waste management sector. Its accessibility would enable small laboratories and institutions to carry out microplastic detection anywhere with an affordable kit and a smartphone. This would facilitate real-time monitoring of plastic pollution and the creation of crowdsourced or user-generated content databases, providing up-to-date information for researchers and policymakers. By facilitating access to this technology across the world, we could identify major pollution hotspots and emerging environmental threats worldwide, which would be helpful in devising timely mitigation strategies. Economically, the use of such a simplified detection method would eliminate the use of specialized laboratory equipment and elaborate infrastructure, reducing costs.

However, while the focus so far has been on microplastics as ecosystem disruptors, these particles have also been found to accumulate in different organisms, including humans (Çobanoğlu, 2021). For humans, the main exposure is through ingestion of contaminated food, inhalation of airborne particles and, to a lesser extent, direct skin contact. In fact, microplastics have been detected in human stool samples. The implications of this have raised the concern even further. LuciPep would be compatible with microplastic screening in human samples. Thus, it would facilitate research on microplastic bioaccumulation and, in the long-term, its impact on human health.

As simplified and accessible as this technology is, we consider its potential limitations. The sensor’s compatibility to certain plastics could limit its effectiveness and narrow its applicability, while technical issues related to luminescence calibration, data handling of users and data accuracy and reliability would have to be assessed and optimized. In the end, the success of this application will partly depend on solving these issues and ensuring broad data sharing and user management, requiring extensive outreach and spreading awareness too. Effective standardization and scalability also play a key role in the promotion and implementation of this application.

References

Bauten, W., Nöth, M., Kurkina, T., Contreras, F., Ji, Y., Desmet, C., ... & Schwaneberg, U. (2023). Plastibodies for multiplexed detection and sorting of microplastic particles in high-throughput. Science of The Total Environment, 860, 160450.

Çobanoğlu, H., Belivermiş, M., Sıkdokur, E., Kılıç, Ö. & Çayır, A. (2021). Genotoxic and cytotoxic effects of polyethylene microplastics on human peripheral blood lymphocytes. Chemosphere, 272, 129805.

Li, J., Liu, H. & Chen, J.P. (2018). Microplastics in freshwater systems: A review on occurrence, environmental effects, and methods for microplastics detection. Water research, 137, 362-374.

Lv, L., Yan, X., Feng, L., Jiang, S., Lu, Z., Xie, H., Sun, S., Chen, J. & Li, C. (2021). Challenge for the detection of microplastics in the environment. Water Environment Research, 93(1), 5-15.

Oh, S., Hur, H., Kim, Y., Shin, S., Woo, H., Choi, J., & Lee, H. H. (2021). Peptide specific nanoplastic detection based on sandwich typed localized surface plasmon resonance. Nanomaterials, 11(11), 2887.

Prata, J. C., da Costa, J. P., Duarte, A. C., & Rocha-Santos, T. (2019). Methods for sampling and detection of microplastics in water and sediment: a critical review. TrAC Trends in Analytical Chemistry, 110, 150-159.

Setälä, O., Granberg, M., Hassellöv, M., Karlsson, T., Lehtiniemi, M., Mattsson, K., Strand, K., Talvitie, J. & Magnusson, K. (2019). Monitoring of microplastics in the marine environment: Changing directions towards quality controlled tailored solutions. Nordic Council of Ministers.

UNEP. (2021). From Pollution to Solution.

Woo, H., Kang, S. H., Kwon, Y., Choi, Y., Kim, J., Ha, D. H., ... & Choi, J. (2022). Sensitive and specific capture of polystyrene and polypropylene microplastics using engineered peptide biosensors. RSC advances, 12(13), 7680-7688.

Yao, Z., Zhang, B. S., & Prescher, J. A. (2018). Advances in bioluminescence imaging: new probes from old recipes. Current opinion in chemical biology, 45, 148-156.

Yee, M., Hii, L., Looi, C.K., Lim, W., Wong, S., Kok, Y., Tan, B., Wong, C. & Leong, C. (2021) Impact of Microplastics and Nanoplastics on Human Health. Nanomaterials, 11(2), 496.

Potential customers and unmet needs

The world has taken a step forward by acknowledging microplastics. However, science needs to continue to move in the right direction by finding a way to obtain comparable data on their detection and quantification. Microplastic identification frequently requires visual classification, which benefits from the use of additional instruments (Lv et al., 2019). A limitation to this is the unsuitability of these methods for the detection of the much smaller nanoplastics, which can result in miscalculations of microplastic content.

Literature points towards the difficulties of using only one method for microplastic and nanoplastic quantification in water samples (Li, Liu & Chen, 2018). So far, the combination of different techniques has been found to significantly improve the study of microplastics as an environmental problem. It also provides more reliable data that can be used for management and policy development. At the same time, though a lot of data on microplastics has been generated, the comparability has been made difficult or impossible because of the use of different methods (Lv et al., 2019).

These limitations point to the immediate need to find a method that is 1) standardized and 2) nanoplastic-sensitive. Current sampling methods often include volume reduction, sample purification and separation (Li, Liu & Chen, 2018), with their due advantages and disadvantages. Thus, our team proposes a new, innovative molecular method that focuses on direct microplastic quantification of a raw, unpurified, sample and that can be developed to include nanoplastic detection.

We address the need for the integration of monitoring protocols for aquatic microplastics. This has been a persistent subject of conversation for many years, but no agreement has been reached on how to achieve it (Nordic Council of Ministers, 2019). For this reason, our project targets important issues that have been raised for microplastic monitoring:

Furthermore, we hope to address the need for cheaper tools that would be more accessible to smaller laboratories around the world. Compared to sophisticated and hyper-specialized tools and machines commonly used to estimate analytes’ concentrations through imaging, the ease and much more immediate access to smartphones could prove an powerful asset for our product.

Use-case scenarios and potential customers

Because of the lack of products that address the previously mentioned problems with microplastic detection, our product has important potential users and a variety of scenarios where it could be a powerful tool. Here, we focus on research and resource management-related scenarios, which can have great impact.

Scenario 1: Environmental research institutes

• The potential customer: research institutions dedicated to environmental studies and water quality assessment.
• Scenario: environmental research institutes often conduct studies on the presence and impact of microplastics in aquatic ecosystems. These organizations require reliable and standardized methods for quantifying microplastics and nanoplastics to support their research initiatives. LuciPep's innovative molecular detection mechanism provides a solution that directly addresses previous diagnostic methods and the limitations. One of LuciPep's fundamental contributions to research would be to increase the accuracy of microplastic quantification data. This directly appeals to the needs of environmental research institutes seeking to generate robust and comparable data for scientific research and policy recommendations.

Scenario 2: Water resource management agencies

• The potential customer: government agencies responsible for managing water resources and ensuring water quality standards.
• Scenario: water resource management agencies are tasked with safeguarding aquatic ecosystems and human health from the impacts of pollution, including microplastics. Reliable methods for monitoring microplastic pollution are essential for these agencies to assess the health of water bodies and develop effective strategies for pollution control. LuciPep's technology offers a standardized and nanoplastic-sensitive approach that can streamline monitoring efforts. More accurate assessments of microplastic contamination in various water sources would enable more efficient and effective regulatory actions and policies to mitigate pollution effectively.

Scenario 3: Small and resource-limited laboratories

• The potential customer: smaller research laboratories and educational institutions with limited resources for sophisticated equipment.
• Scenario: many smaller laboratories, particularly those in developing regions or educational institutions, face challenges when it comes to accessing and affording advanced equipment for complex analyses. These laboratories often lack the resources to invest in high-cost instrumentation, such as the one that could be required for bioluminescence imaging and quantification. LuciPep's innovative smartphone-based bioluminescence detection method would present an ideal solution for these laboratories. By offering a cost-effective and user-friendly detection approach, LuciPep enables smaller laboratories to overcome financial and technical barriers. The relative simplicity of using a smartphone (much more accessible, immediate and widely spread than most laboratory equipment) for the direct and accurate quantification of microplastics and nanoplastics makes it an accessible option for labs with limited resources. This opens up opportunities for educational institutions and research laboratories in remote or resource-constrained areas to contribute to the global understanding of microplastic pollution. LuciPep's collaboration with such labs could empower researchers and students to engage in impactful environmental research, providing them with a practical tool to measure microplastic contamination and contribute to the broader scientific knowledge about plastic pollution.

Collaborations

• Research universities and institutes with expertise in molecular biology, environmental science, and water quality assessment can be valuable collaborators. Collaborative efforts can help refine and validate LuciPep's technology, ensuring its accuracy and reliability across different environments and conditions.
• Environmental non-governmental organizations (NGOs) focused on plastic pollution and conservation can provide real-world insight and access to water samples from diverse locations. Collaborating with these organizations can facilitate field testing and validation of LuciPep's method, enhancing its applicability and demonstrating its impact.
• Companies specializing in biotechnology, molecular diagnostics and instrumentation can offer technical expertise and resources to further develop and optimize LuciPep's detection method. Collaborating with established biotech companies can accelerate the translation of the technology from concept to a market-ready product.
• Partnerships with government agencies responsible for environmental protection and public health can provide regulatory guidance and funding opportunities for research and development. Collaborating with these agencies can help ensure that LuciPep's technology aligns with regulatory standards and addresses pressing environmental concerns.
• Analytical laboratories with experience in microplastic analysis can facilitate rigorous testing and validation of LuciPep's method against established techniques. This collaboration can lead to the adoption of LuciPep's technology as a reliable and standardized tool for microplastic detection and quantification.

Feasibility

Microplastic contamination poses a severe environmental challenge with significant implications for aquatic ecosystems and public health. Current estimates project that plastic waste entering aquatic ecosystems may reach between 23 and 37 million tons annually by 2040, emphasizing the urgency of effective monitoring and management (UNEP, 2021). However, the scale of microplastics under 1 mm, coupled with the lack of routine monitoring, presents a substantial obstacle for accurate quantification (Prata et al., 2019).

Current microplastic detection methods encompass a range of analytical approaches that employ microscopy and mass spectrometry techniques. These methods have important limitations. For instance, fluorescence microscopy, commonly used in initial inspections with staining dyes like Nile Red, offers limited insight into chemical composition and unreliable quantification (Bauten et al., 2023). On the other hand, spectroscopy methods, such as Fourier-transform infrared spectroscopy (FTIR) and Raman spectroscopy, along with mass spectrometry, provide chemical composition data but often involve time-consuming processes and specialized equipment (Prata et al., 2021). Consequently, a universal protocol for microplastic detection remains elusive.

Our innovative microplastic biosensor, LuciPep, addresses these challenges with a focus on feasibility. LuciPep leverages the specificity of microplastic binding peptides (MBPs) to bind to different microplastic types, enabling targeted detection (Woo et al., 2022). Notably, LuciPep incorporates NanoLuc luciferase, a bioluminescent enzyme, as a biosensor. This enzyme's oxidation of furimazine produces detectable light, eliminating the need for external light sources and background emissions (Yao et al., 2018). As a result, LuciPep offers highly accurate quantification, enhanced specificity and superior sensitivity compared to conventional methods.

Moreover, LuciPep's integration with smartphones for data analysis ensures accessibility and convenience, reducing the reliance on costly and specialized equipment. Researchers and water treatment operators can utilize their smartphones to detect and quantify microplastics conveniently. The amalgamation of accurate quantification, specificity, sensitivity and smartphone compatibility positions LuciPep as a feasible and transformative tool for microplastic detection.

Viability

Traditional microplastic detection methods often demand expensive equipment, such as fluorescent microscopes and spectrometers, rendering them financially burdensome for many users. However, ensuring the viability of microplastic detection methods requires affordable and accessible solutions. LuciPep addresses this issue by circumventing the need for high-end, expensive equipment. First of all, the biosensor itself can be produced cost-effectively, and its compact design enhances portability. Field researchers and water treatment plant operators can conduct tests directly in the field, eliminating the need for dedicated laboratory facilities.

Furthermore, LuciPep's compatibility with smartphones provides a level of accessibility that traditional methods cannot match. By using the smartphone's camera and basic image analysis software, it provides a cost-effective and widely available solution that extends its accessibility to smaller laboratories and resource-limited institutions. This democratizes microplastic detection, enabling users to carry out tests conveniently and efficiently. As a result, LuciPep emerges as a viable, cost-effective and accessible alternative for microplastic detection.

Scalability

The scalability of LuciPep depends on its integration into existing practices and its widespread adoption within the scientific community and environmental monitoring agencies. Here, a critical factor will be facilitating a smooth assimilation of the method by existing microplastic detection protocols. Because of LuciPep’s previously discussed accessibility and affordability, researchers and water treatment facilities can readily incorporate LuciPep into their workflow with minimal adjustments. Collaboration with research institutions, environmental agencies and analytical laboratories can provide real-world testing and validation, boosting confidence in the technology's reliability and fostering its adoption.

On the same line, regulatory support and endorsements from government agencies responsible for environmental protection are pivotal for scalability. Collaborating with these agencies can help establish LuciPep as a trusted and recommended tool for microplastic monitoring. To further reinforce its adoption, awareness campaigns and training programs to educate users and enhance competence and familiarity with the technology would be advisable.

By ensuring compatibility with current practices, fostering collaborations, seeking regulatory support and promoting user awareness, LuciPep can be effectively implemented and adopted on a large scale, contributing to comprehensive microplastic monitoring efforts.

Inventivity

LuciPep's novelty lies in its innovative combination of molecular biology, bioluminescence and smartphone technology to address the challenges of microplastic detection. It breaks new ground in several aspects:

1. LuciPep capitalizes on the recent development of microplastic binding peptides (MBPs) for high-specificity microplastic detection (Woo et al., 2022). These short amino acid sequences offer the unique ability to target specific microplastic types, departing from traditional methods.
2. LuciPep integrates NanoLuc luciferase as a bioluminescent biosensor. This enzyme generates light without external stimulation, eliminating background emissions and enhancing sensitivity (Yao et al., 2018). Such bioluminescent detection is an innovative approach to microplastic quantification.
3. LuciPep's compatibility with smartphones for data analysis adds a new layer of accessibility and convenience for users and eliminates the requirement for specialized and costly equipment.
4. LuciPep's sensitivity extends to the detection of nanoplastics, an area that has remained challenging within the field (Oh et al., 2021). Its ability to detect particles below 1 µm in size is a unique feature, addressing an unmet need in microplastic monitoring.

SWOT analysis

In the process of creating LuciPep, we dedicated a significant amount of time to defining our milestones and the strategies to achieve them. We started by conducting a SWOT analysis to evaluate both internal and external factors, particularly the potential risks they could pose to our company. After identifying the strengths and potential risks, we developed a roadmap that identified the key areas of development and their corresponding timelines. Additionally, we identified stakeholders and the necessary capabilities required to turn these milestones into reality. We assessed our product’s strengths and opportunities, as well as the weaknesses and threats we need to address by conducting a SWOT analysis.

Figure 1: SWOT analysis for LuciPep.

LuciPep offers an user-friendly experience, removing barriers and reducing costs. The integration of smartphone-based analysis, without the need for built-in infrastructure, extends accessibility to anyone with basic molecular biology skills and a smartphone. This inclusivity aligns with LuciPep's vision of making accurate detection of microplastics universally attainable. The adaptability of LuciPep's technology is yet another distinct strength. It can be swiftly adapted to detect different types of microplastics, ensuring versatility and market relevance. Furthermore, its potential to evolve into a detection and purification system adds value and diversity to its applications.

An opportunity for LuciPep is entering the market as a microplastic detection technology for blood samples. This represents an exciting frontier in environmental and health research. By leveraging the same innovative approach that enables accurate and user-friendly detection of microplastics in water, LuciPep could open new avenues for studying the presence of microplastics within the human body. This groundbreaking capability could facilitate research into the health implications of microplastic exposure, offering valuable insights into their potential effects on human physiology. Thus, LuciPep's adaptability enables it to contribute not only to environmental conservation but also to advancements in medical and public health research, ultimately broadening the scope of its positive impact on our world.

There are also external factors, opportunities, that will help LuciPep to grow as a successful company and deliver its product to end users. The global awareness surrounding the issue of microplastic pollution in our waters continues to grow. This burgeoning awareness translates into an escalating demand for microplastic detection solutions. LuciPep is ready to meet this demand by providing innovative solutions to address the urgent problem of water pollution. Collaboration and endorsement opportunities with governmental and environmental agencies represent a promising avenue. By aligning with these stakeholders, LuciPep can obtain invaluable support and resources that enhance its credibility and reach. Also, the possibility of collaborating with research institutions, academic research groups, environmental organizations, and water management authorities presents a remarkable opportunity. These partnerships not only improve the credibility of LuciPep's product but also foster synergistic research and development efforts, allowing to further enhance the product offered by LuciPep.

Yet, developing LuciPep into a successful company presents both internal and external risks that require careful consideration. One of our main weaknesses lies in the yet unknown accuracy of our product's detection of microplastics in real water samples. If LuciPep's accuracy lags behind that of competitors, it could significantly undermine its credibility. Additionally, the detection of microplastics through our kit depends on the quality of phone cameras, which varies across different brands and models. However, to overcome this problem, the smartphone application will be adapted to work with consistent reliability across imaging hardware variations.

On the other hand, LuciPep might face external threats. The microplastic detection field is rapidly evolving, with potential competitors constantly innovating to produce more feasible, accurate and user-friendly solutions. This competitive landscape poses a significant risk, as we must continually strive to stay ahead in terms of innovation and product quality. Thus, we face the challenge of keeping our product up to date in the face of rapid advancements in biotechnology and sensor technology. This requires a long-term commitment to research and development, requiring appropriate financing and resources.

These identified weaknesses and external threats underscore the need for a comprehensive risk management strategy. Embracing rigorous quality control, continuous innovation, adaptability and regulatory diligence will be key to overcoming these challenges and making LuciPep a successful company. Furthermore, conducting extensive field research is essential to refine our product and adapt it to real-world conditions.

LuciPep’ microplastic detection technology can be adapted to analyzing blood samples in the future, as an exciting frontier in environmental and health research. This groundbreaking capability could facilitate research into the health implications of microplastic exposure, offering valuable insights into their potential effects on human health and physiology. Hence, the adaptability of LuciPep's technology underscores its versatility, enabling it to contribute not only to environmental conservation but also to advancements in medical and public health research, ultimately broadening the scope of its positive impact on our world.

Milestones and timelines

The current stage of LuciPep is dependent on further product development, business establishment and commercialization. We have identified milestones in the research and development of our kit and smartphone app, kit manufacturing, business and financial planning and regulatory compliance process. We estimate that our kit will be commercially launched within the next 3 years. Our roadmap, which outlines the milestones we aim to achieve and their respective timelines, can be found in the Gantt chart presented below.

Figure 2: Development Roadmap of LuciPep, represented as a Gantt chart.

Stakeholders and required skills

Next, we have identified the stakeholders involved in these milestones and classified them based on their potential impact and interest in LuciPep. This should help to better understand our organization’s interaction with them.

Figure 3: Stakeholder map.

Click here to view the table with stakeholders and the resources required to achieve our milestones.

We strongly believe that collaborations with biotech incubators such as Karolinska Institutet Innovations AB and KTH Innovations will provide the foundation for prototype development and technical validation. These partnerships will empower us to create tangible prototypes. While developing our prototype, we can leverage the services of the Protein Science Facility (PSF) at Karolinska Institutet, taking advantage of the cost-effective resources it offers to KI students and academics. Additionally, partnering with prototype development companies like Prototyp Stockholm will enable us to manufacture both our prototype and our user-friendly app. After successful prototype testing, we can collaborate with Nordic BioSite to scale up our recombinant protein production.

To make these milestones achievable, we also need to consider our funding opportunities. Partnering with biotech incubators will enable us to apply to pre-seed fundings. KTH Innovation manages the Vinnova funding program, VFT-1, for startups in need of early-stage funding. They offer up to 300 000 SEK ($27.128 in October 2023). Following prototype development, our goal is to secure seed funding from angel investors, incubators and early-stage venture capital firms specializing in early-stage startups. In this phase, we aim to secure funding from Almi Invest GreenTech Fund, a Swedish business developer with €65,000,000.00 ($68,813,434 in October 2023) in managed funds, dedicated to funding early-stage Swedish startups. In a more advanced stage, we aim to apply to a series of funding rounds from venture capital firms. Nevertheless, we acknowledge that there may be a need for additional support from bank loans and contributions from family and friends throughout this journey.