Contribution | TJI-Seoul

Introduction

From the very beginning of our project, our team was committed to making sure that our work was not only scientifically sound, but also socially relevant and meaningful to real-world needs. To achieve this, we integrated Human Practices into the core of our design process. Before finalizing our experimental plan, we conducted public awareness surveys to better understand how the general public perceives cancer immunotherapy and protein engineering. These results revealed knowledge gaps and helped us tailor our education efforts, but they also gave us insight into how to communicate our project in a way that is both accessible and scientifically rigorous.

In parallel, we interviewed experts in computational biology, immunology, and protein engineering. Their feedback guided the technical aspects of our design: for example, highlighting both the strengths and limitations of in silico prediction tools, and reminding us of the importance of experimental validation. By listening to these voices early on, we were able to refine our approach to engineering MHC Class I proteins in a way that addresses scientific challenges while considering practical applications in medicine. This combination of community input and expert advice allowed us to design experiments that are not only academically interesting but also aligned with actual demands in the medical and scientific field.

This vision led to the development of the HLA Engineering Toolkit, a standardized, modular system for expressing, refolding, and analyzing human MHC Class I proteins (HLA-A*02:01 and β2-microglobulin) in E. coli. By combining computational protein design, experimental validation, and educational outreach, our project bridges three key aspects of iGEM: scientific innovation, reproducibility, and accessibility. The resulting toolkit is not only an original contribution to the iGEM Parts Registry but also a valuable educational and research resource for future teams working in immunology, protein engineering, or computational biology.

Parts & Parts Collection Contribution

In the laboratory, we established a protocol pipeline for improving and enhancing protein function, using the MHC Class I protein as our test case. This work required us to design and construct plasmids encoding both wild-type and mutant variants of HLA-A*02:01 heavy chains, as well as β2-microglobulin (B2M).

Our major technical contribution is the creation of a four-part plasmid system designed for modular, reproducible engineering of MHC Class I proteins. The toolkit includes: HLA Engineering Toolkit (New Parts Collection): c64ee9f8-f6ec-4ff4-9da1-38ee16033a42

Wild-Type HLA-A*02:01 Heavy Chain (BBa_25LBQEGY) — the baseline construct for benchmarking folding, stability, and peptide-binding affinity.
Mutant 1 (W167A) Heavy Chain Variant (BBa_25SNBFGX) — a single-point mutation designed via AI modeling (DiffDock) to alter the peptide-binding groove’s flexibility and enhance antigen recognition.
Mutant 2 (Y7A/Y99A/Y159A/Y171A) Heavy Chain Variant (BBa_25SCBFYY) — a quadruple mutant targeting multiple tyrosine residues to explore cooperative structural changes in peptide accommodation.
Human β2-Microglobulin (BBa_25GNHXKA) — an essential non-tagged light chain required for proper folding and complex formation with the heavy chains.

The detailed step-by-step procedures for each experiment can be found on our Experiments page, where they are described in a way that can be reproduced by other teams.

Beyond technical details, our contributions establish a generalizable workflow for enhancing protein functions through engineering. Although our immediate application is cancer therapy, this pipeline can be extended far beyond oncology:

Contribution to the iGEM Registry

The same strategy of improving protein stability and binding affinity can be applied to other therapeutic proteins, such as enzymes used for treating metabolic disorders, antibodies for autoimmune diseases, or cytokines used in immune regulation. By optimizing folding, binding affinity, or receptor recognition, our approach could provide a way to increase the safety and effectiveness of biologic drugs.

Applications in Other Therapeutic Areas

Applications in Industrial Biotechnology

Protein engineering is not limited to medicine. Many industrial processes rely on enzymes as catalysts. By adapting our pipeline for enzyme optimization, industries could develop more stable and efficient biocatalysts for use in manufacturing, energy, or environmental technology. For instance, enzymes designed to resist high temperatures could make industrial processes more sustainable and cost-efficient. Similarly, agricultural biotechnology could benefit from engineered proteins that enhance plant immunity against pathogens, contributing to improved food security.

In this way, our wet lab contributions go beyond immediate research---they provide a replicable system that other teams and researchers can adapt for multiple contexts, from drug development to industrial innovation. Thus, our wet lab work is not a one-off demonstration but a reusable framework for enhancing protein function, which can serve future iGEM teams and researchers.

Scientific Contribution and Significance to the iGEM Registry

The HLA Engineering Toolkit represents both a scientific advancement and a significant addition to the iGEM Parts Registry. Each plasmid contributes individually as a functional genetic part, but their true value lies in how they work together as an integrated collection. The toolkit offers a standardized, modular framework for studying MHC Class I biology — something rarely seen in synthetic biology projects.

From a scientific perspective, the toolkit demonstrates that complex human immune proteins can be rationally engineered and functionally reconstructed in bacterial systems. This opens new opportunities for synthetic immunology research by providing an accessible method to study antigen presentation, folding stability, and mutation effects outside of mammalian cell culture.

By establishing a consistent experimental baseline through the wild-type HLA construct, our mutants could be evaluated objectively — allowing accurate comparison of how each mutation alters folding or peptide binding. The inclusion of β2-microglobulin ensures that these systems mimic real physiological assembly, making the results biologically meaningful.

From an iGEM contribution standpoint, our work fulfills several key goals of the competition:

It provides a complete, ready-to-use parts collection, rather than isolated DNA sequences.
It offers a replicable experimental system that other teams can use to study immune-related proteins.
It bridges computational prediction and wet-lab validation, helping to connect AI-driven protein design with real-world biochemistry.
It expands the Parts Registry into new territory — introducing standardized resources for immunological and therapeutic protein research.

The HLA Engineering Toolkit is therefore more than a set of constructs — it is a framework for understanding and engineering human immune proteins. By registering these parts and documenting the full process of design, measurement, and validation, we provide future teams with a strong foundation for both research and innovation in this field.

Education Contribution

Education was a cornerstone of our project, as we believe scientific advances should go hand in hand with raising public awareness and engagement. To design our outreach efforts, we first conducted a public awareness survey on cancer, immunology, and synthetic biology. The survey revealed that while many people had a general understanding of cancer, there was limited knowledge of how the immune system detects abnormal cells and how therapies like immunotherapy work.

Based on these results, we developed an educational presentation tailored to the public's current level of awareness. Instead of overwhelming audiences with technical jargon, we built our content step by step: starting with the basics of the immune system, then explaining the role of MHC proteins, and finally showing how protein engineering can lead to improved cancer treatments. We also included interactive visuals and case examples to make complex concepts accessible and engaging.

The purpose of this education initiative was twofold:

To bridge the knowledge gap by making cancer immunology and synthetic biology understandable to high school students and general audiences.
To create long-term impact by fostering interest in biotechnology and showing how synthetic biology can contribute to solving major health challenges.

By tailoring the presentation to the survey results, we ensured that our educational efforts were not generic but directly responsive to public needs. We believe this model can inspire other iGEM teams to integrate feedback-driven education into their own outreach strategies.

Summary of Contributions

In summary, our contribution to iGEM combines technical innovation with educational impact. Through the creation of the HLA Engineering Toolkit, we provide a reproducible, open-source platform for engineering and analyzing MHC Class I proteins — a major step toward establishing synthetic immunology within the iGEM framework.

Our system bridges computational modeling and laboratory experimentation, offering both a scientific resource and a practical workflow for future teams. Beyond the lab, our outreach initiatives ensure that the science behind our project reaches broader audiences, connecting biotechnology with public understanding.

Together, these contributions embody the core spirit of iGEM — advancing synthetic biology through collaboration, accessibility, and creativity. The HLA Engineering Toolkit stands as a foundation that future teams can build upon to explore new frontiers in immune system engineering and biomedical innovation.