Protein Engineering · Computational Biology

Engineering IL-10 for improved thermostability.

A protein engineering project focused on identifying IL-10 variants with improved predicted thermostability while maintaining structural compatibility with IL-10 receptor interactions.

01 · THE PROBLEM

Why engineer IL-10?

Interleukin-10 (IL-10) is an immunoregulatory cytokine with therapeutic potential for inflammatory diseases including inflammatory bowel disease (IBD).

One challenge in developing protein therapeutics is balancing functional activity with physical stability. This project explored whether computational protein design could identify IL-10 variants predicted to have improved thermostability while preserving the structural features required for receptor binding.

02 · DESIGN STRATEGY

From mutation design to structural evaluation.

I worked with a protein design team to combine multiple computational approaches for evaluating candidate IL-10 mutations.

IL-10
→
Candidate Mutations
→
ThermoMPNN
→
ESM
→
AlphaFold3
→
Variant Selection
03 · COMPUTATIONAL DESIGN

Screening candidate mutations.

Computational protein design was used to narrow the search space before moving toward experimental construction. ThermoMPNN was used to evaluate candidate mutations based on predicted effects on protein stability.

ThermoMPNN heatmap for IL-10 mutation design
ThermoMPNN heatmap showing predicted stability effects across candidate IL-10 mutations.
01 · ThermoMPNN

Stability prediction

Screened candidate mutations based on predicted effects on protein thermostability.

02 · ESM

Sequence-level evaluation

Used protein language model-based analysis as an additional computational filter for candidate designs.

03 · AlphaFold3

Structural evaluation

Structural modeling was used to examine selected variants and assess their predicted conformations.

04 · SELECTED VARIANTS

Two computationally selected variants.

After computational screening, two IL-10 variants were selected for further evaluation. The variants were examined using structure prediction to assess whether the proposed mutations were compatible with the overall IL-10 fold.

AlphaFold structure of IL-10 Mutation Variant 1
Selected Design

Mutation Variant 1

AlphaFold-based structural visualization of the first selected IL-10 mutation variant.

AlphaFold structure of IL-10 Mutation Variant 2
Selected Design

Mutation Variant 2

AlphaFold-based structural visualization of the second selected IL-10 mutation variant.

05 · EXPERIMENTAL DESIGN

Iterating toward a production plasmid.

Computational design was followed by iterative plasmid design. The construct evolved through multiple design iterations before reaching a finalized production configuration.

First iteration of IL-10 plasmid design
Iteration 01

First plasmid design

Initial construct design developed for the experimental stage of the project.

Second iteration of IL-10 plasmid design
Iteration 02

Second plasmid design

Revised construct following the first design iteration.

Finalized IL-10 production plasmid
Final Design

Production plasmid

Finalized construct prepared for downstream experimental production.

Public portfolio note: detailed sequence information, unpublished constructs, or proprietary experimental information should only be displayed if approved for public release.
06 · CONSTRUCT VISUALIZATION

Visualizing the engineered construct.

ApE was used to visualize and inspect the plasmid architecture during the construct design process.

ApE visualization of IL-10 plasmid
ApE visualization used during IL-10 plasmid design and inspection.
07 · MY ROLE

Protein design team leadership.

I led a 20+ member protein design team working on the IL-10 engineering project. My work involved coordinating computational protein design, evaluating candidate mutations, interpreting structural predictions, and helping connect computational results to downstream experimental design.

20+
Protein design team members
2
Selected IL-10 variants
3
Major computational approaches
08 · DOCUMENTATION

Project presentation.

The work was also presented through scientific poster communication, providing a broader overview of the project, engineering strategy, and results.

View BMES Poster →
This project was completed as part of a larger protein engineering effort and contributed to an iGEM Gold Medal and Inclusivity Award.