Periodic Reporting for period 2 - EVOdrops (directed EVOlution in DROPS)
Periodo di rendicontazione: 2020-10-01 al 2023-09-30
The bio-economy is poised for substantial growth, with projections indicating that by 2030, biotechnological production will contribute significantly to various sectors. Estimates suggest a substantial presence, such as 35% in chemical and industrial products, 80% in pharmaceutical and diagnostic production, and 50% in agricultural output. Industries, continually seeking process and product enhancements, face challenges in natural evolution's slow pace. Natural processes, including Darwinian evolution, optimize proteins in response to environmental pressures over billions of years. Enzymes, as nature's catalysts, play vital roles in diverse cellular reactions. While some natural enzymes hold industrial promise, relying on natural evolution for their improvement has drawbacks: limited control over selection pressures and inability to enhance non-natural systems. Directed evolution, mimicking nature's principles in the laboratory, addresses these limitations. Unlike the lengthy natural evolution, directed evolution combines rapid genetic diversity generation, ultra-high throughput screening, and selection for efficient industrial processes. Microfluidics, a versatile technology, integrates into industrial processes, employing droplets as micro-reactors in miniaturized, automated assays. This enables ultra-high-throughput screening, reducing reagent costs and facilitating experiments impossible with other techniques. The microfluidic market has witnessed impressive growth, driven by start-ups and industrial giants entering the sector. The technology's potential applications in diagnostics, healthcare, and bio-resource transformation contribute to its expanding market. In a later project phase, Artificial Intelligence (AI) emerged as a crucial element. Recognizing AI's potential in microfluidics and directed evolution, the EVOdrops project incorporated it. AI, applied in health and medical fields for early diagnosis and personalized medicine, extends its utility to other industrially important molecules. The convergence of AI with micro-/acoustofluidics expands the EVOdrops portfolio, emphasizing the interdisciplinary approach for enhanced outcomes.
This integration aligns with the evolving landscape of technological development, with AI gaining prominence during the pandemic. The recognition of AI's significant potential prompted its inclusion in the EVOdrops program, showcasing a dynamic response to technological advancements.
This integration aligns with the evolving landscape of technological development, with AI gaining prominence during the pandemic. The recognition of AI's significant potential prompted its inclusion in the EVOdrops program, showcasing a dynamic response to technological advancements.
EVOdrops merges microfluidics and biochemical methods to engineer proteins and systems for industrial or therapeutic use. It trains a diverse group of Early-Stage Researchers (ESRs) across biology, biotechnology, microfluidics, and microtechnology engineering, aiming to transcend single-team or institutional constraints in exploring directed evolution processes for protein engineering. ESRs adopt a global, multi-disciplinary approach, utilizing key enabling technologies for high-throughput biological analysis. The overarching objective is to optimize processes for ultra-high throughput, improved sensitivity, and system reliability in selecting variants of interest. During Period 1, the project established a Supervisory Board, fostering collaboration, hiring and training ESRs, and creating multidisciplinary ESR supervision panels. Despite pandemic challenges, six secondments occurred, and initial scientific results on library manipulations and microfluidic integration were obtained. Due to the pandemic, fewer secondments occurred, emphasizing Workshop program completion and inter-project coherence. ESRs prioritized becoming leaders in directed evolution and microfluidics, with the first milestone being a doctoral qualification. The majority have submitted their PhD theses, and four have obtained their PhD qualifications. Anticipating 12 out of 13 ESRs will submit a thesis, one replacement ESR lacked sufficient research time. Dissemination involves a website, Twitter account, and an industrial newsletter initially managed by PIs, the network, and the scientific liaison manager. The second period planned increased ESR involvement in platform design, but pandemic restrictions shifted focus to research. ESRs participated in live-streamed video presentations at the 2012 Explorathon, showcasing directed evolution and microfluidics. ESR4's group excelled in the international Directed Evolution Competition, winning a gold medal.
In the consortium EVOdrops, a central impact goal is equipping Early-Stage Researchers (ESRs) with tools and skills for successful, long-term career planning, offering training for potential positions in both industry and academia. Leveraging industrial partners, beneficiaries, and commercial companies, EVOdrops broadens ESRs' perspectives on career options through talks and site visits. Talks during Workshop 1 (WS1) featured insights from BioMillenia and Thermo Fisher Scientific Baltics on strategy and working environments, while Workshop 2 (WS2) had IBIDI discussing the challenges in an SME. In the second project period, emphasis was placed on commercial company involvement with site visits to Dolomite Micro, Dolomite Bio, Sphere Fluidics, Treefrog, and Emulseo. The challenging pandemic situation necessitated adjustments, leading to postponed workshops. Despite restrictions, transferable skills training, including communication, public engagement, academic writing, and entrepreneurship awareness sessions, was incorporated into all workshops. Positive feedback from ESRs prompted the organization of 'career talks' to further enrich their understanding of potential career paths. On the scientific front, EVOdrops aims to advance the state of the art. The program has introduced new gene libraries, methods, and protocols for directed evolution, along with innovative chip designs for precise droplet production, manipulation, and sorting. The establishment of enhanced production facilities has expanded capabilities. During research, new microbial species were isolated. Notable scientific achievements include the development of a micro-fluidic platform capable of evolving proteins toxic to cells. New protein expression methods were devised for single DNA particles with multiple copies of the same gene. Sorting devices were improved by incorporating multiple detectors, resulting in an >800-fold enrichment in a single round. Advanced droplet generation covering three orders of magnitude in size and volume was achieved, contributing to increased sorting capabilities. Furthermore, the consortium achieved significant progress in droplet generation, covering size and volume ranges across three orders of magnitude. This breakthrough enhances the versatility and applicability of the technology, making it a valuable tool for a wide range of applications. Other groups reported identifying previously uncultured microbial species. ETH introduced a double selection system and FACS sorting, simplifying the sorting of population fractions. TUM developed a new riboswitch for directed evolution and plans to explore novel riboswitches from aptamers. Additionally, TUM and CNRS Bordeaux established a pico-injection system, ensuring controlled component delivery into droplets. These scientific advancements contribute to the broader goals of EVOdrops, pushing the boundaries of directed evolution and microfluidics.