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Updated: Feb 10, 2026
21:08
Engineering Cell-permeable Protein
Published on: December 28, 2009
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Fn3 proteins engineered to recognize tumor biomarker mesothelin internalize upon binding
Allison R Sirois1,2, Daniela A Deny3, Samantha R Baierl2
1Molecular and Cellular Biology Program, University of Massachusetts, Amherst, Amherst, Massachusetts, United States of America.
Plos One
|May 9, 2018
Summary
Researchers engineered non-antibody proteins called fibronectin type III domain (Fn3) variants to target mesothelin, a protein overexpressed in many cancers. These engineered Fn3 variants show promise for cancer diagnostics and therapeutics.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Mesothelin is a cell surface protein overexpressed in various cancers, promoting tumor progression and metastasis.
- Targeting mesothelin holds potential for cancer diagnostics and therapeutics, but approved molecules are lacking.
- Non-antibody protein scaffolds offer advantages over antibodies for certain clinical applications like molecular imaging and drug delivery.
Purpose of the Study:
- To engineer non-antibody protein variants that specifically bind to mesothelin with high affinity.
- To evaluate the potential of these engineered proteins for targeted cancer diagnostics and therapeutics.
Main Methods:
- Utilized directed evolution and yeast surface display to engineer fibronectin type III domain (Fn3) variants.
- Produced and purified lead engineered Fn3 variants from bacterial culture.
- Assessed binding specificity to mesothelin on human cancer cell lines and evaluated cellular internalization.
Main Results:
- Successfully engineered high-affinity Fn3 variants that specifically bind to mesothelin.
- Engineered Fn3 proteins were produced in high yield and purified.
- Upon binding to mesothelin, the Fn3 variants internalized and co-localized to early endosomes in cancer cells.
Conclusions:
- Demonstrated the successful engineering of non-antibody proteins (Fn3 variants) to target the cancer biomarker mesothelin.
- These findings validate the use of engineered protein scaffolds for targeting mesothelin.
- Encourages further development of these engineered variants for clinical applications in targeted cancer diagnostics and therapeutics.
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