RESULTS

PCR Proof of Concept • Western Blot Refinement • Pre-commercial Assessment

Experimental Progress Overview

During our project, we progressed through Bacmid design, amplification, and transfection. Preliminary PCR checks provided initial evidence that the Bacmid successfully entered the baculovirus system, suggesting that our genetic design had the potential to reach host cells as intended.

While these early results are promising, we continued to validate the expression more rigorously through additional analytical methods.

Protein Expression Validation

As we conducted Western blotting to confirm gene expression, we obtained bands with limited clarity, which we attributed to procedural aspects that affected protein sample quality.

Systematic Process Improvement

We spent time refining the procedure and prepared protein samples again, demonstrating our commitment to methodological rigor and systematic troubleshooting.

Phenotypic Testing & Genetic Stability Assessment

We attempted phenotypic testing by measuring the overnight survival rate of frozen cells using hemocytometer counts. Analysis revealed that cells undergoing multiple passages showed variable performance.

Careful investigation identified that these cells risked the loss of inserted DNA—a finding that highlights the importance of genetic stability considerations for long-term cell culture applications.

Technical Validation & Key Insights

Proof of Concept Established: Preliminary PCR indicates successful Bacmid integration, while phenotypic testing confirms the potential of tardigrade proteins for cell protection during storage.

Methodological Refinement Pathway: Western blotting identified procedural optimization opportunities, leading to immediate protocol improvements.

Genetic Stability Challenge Identified: Multiple passages revealed insert retention challenges, informing our risk assessment for commercialization.

Commercial Viability Demonstrated: The core concept of using tardigrade protective proteins shows promise, though further refinement is needed before market readiness.

Optimization Strategy & Commercial Pathway

Rather than viewing these challenges as setbacks, we treat them as key lessons in the engineering cycle. Based on existing achievements, we have planned our next steps with commercial success in mind.

Strategic Development Pathway

Further investigation and targeted improvement of the transfection step to address DNA retention challenges

Continued refinement of protein analysis methodologies based on lessons learned

Systematic approach to bridge the gap between proof-of-concept and commercial readiness

This optimization pathway is crucial for our product to be successful on the market

Entrepreneurial Perspective

From an entrepreneurial perspective, these experiences are valuable—they highlight real-world bottlenecks such as unexpected procedural flaws and inconsistent gene expressions that must be addressed for successful commercialization.

By documenting these experiences transparently, we are not only refining our own process but also providing guidance for future teams who may want to scale tardigrade-protein-based solutions.

Future Outlook

We are excited to continue refining our workflow and anticipate that the next round of experiments will provide clearer outcomes.

These ongoing efforts illustrate the potential of tardigrade proteins to improve cell survival and reduce research costs, demonstrating both scientific curiosity and practical foresight for commercial applications.