Periodic Reporting for period 1 - trEM4all (Making time-resolved cryo-EM available for the community of structural biologists: validate, improve, derisk)
Berichtszeitraum: 2024-02-01 bis 2025-07-31
Experimental determination of protein structures is central to molecular biology and drug development. Cryogenic electron microscopy (cryo-EM) has revolutionized structural biology by enabling rapid structure determination of proteins vitrified in solution. However, current cryo-EM sample preparation methods limit the observation of dynamic processes and the use of low-yield or native samples.
To address these limitations, we developed two complementary microfluidics-based technologies:
- Time-resolved cryo-EM (trEM): enabling structural snapshots of proteins in action.
- Microfluidics-based Isolation for Sample Optimization (MISO): facilitating cryo-EM sample preparation from low-yield or native sources.
Both technologies were at TRL2 at project start and lacked commercial equivalents. The trEM4all project aimed to validate, improve, and derisk these innovations, exploring their broader dissemination and commercialization potential.
To address these limitations, we developed two complementary microfluidics-based technologies:
- Time-resolved cryo-EM (trEM): enabling structural snapshots of proteins in action.
- Microfluidics-based Isolation for Sample Optimization (MISO): facilitating cryo-EM sample preparation from low-yield or native sources.
Both technologies were at TRL2 at project start and lacked commercial equivalents. The trEM4all project aimed to validate, improve, and derisk these innovations, exploring their broader dissemination and commercialization potential.
- trEM Technology: Validated on two new protein classes, including membrane proteins. A new chip design was developed to prevent dye-induced protein damage, enabling high-resolution reconstructions of fragile complexes. A patent was filed for this improved design.
- MISO Technology: Originally developed under an ERC-CoG grant, MISO was integrated into trEM4all for business analysis and scalability studies. It reached TRL4 and attracted strong interest from pharmaceutical companies for its potential to streamline cryo-EM workflows.
- Scalability: Transitioned microfluidic chips from PDMS to thermoplastics using hot embossing, demonstrating feasibility for industrial production of both trEM and MISO chips.
- Commercial Interest: Engaged with cryo-EM manufacturers and pharma companies. While trEM was seen as promising but early-stage, MISO received strong interest for near-term applications. A roadmap for MISO MVP development was created, and additional funding secured to support its transition.
- MISO Technology: Originally developed under an ERC-CoG grant, MISO was integrated into trEM4all for business analysis and scalability studies. It reached TRL4 and attracted strong interest from pharmaceutical companies for its potential to streamline cryo-EM workflows.
- Scalability: Transitioned microfluidic chips from PDMS to thermoplastics using hot embossing, demonstrating feasibility for industrial production of both trEM and MISO chips.
- Commercial Interest: Engaged with cryo-EM manufacturers and pharma companies. While trEM was seen as promising but early-stage, MISO received strong interest for near-term applications. A roadmap for MISO MVP development was created, and additional funding secured to support its transition.
- trEM: Achieved TRL3 with validated applications, improved chip design, and dedicated cryo-EM grid concepts (cGrids).
- MISO: Advanced to TRL4 with validated use cases and strong market traction. A spin-off roadmap and MVP development plan were created.
- Technology Transfer Needs: Further chip refinement, external lab testing, and demo device development are planned. Industrial partnerships and public funding are being pursued to reach TRL6.
- MISO: Advanced to TRL4 with validated use cases and strong market traction. A spin-off roadmap and MVP development plan were created.
- Technology Transfer Needs: Further chip refinement, external lab testing, and demo device development are planned. Industrial partnerships and public funding are being pursued to reach TRL6.