Target-Binding Proteins Based on the 10th Human Fibronectin Type III Domain (10Fn3)
Abstract
We describe concepts and methods for generating a family of engineered target-binding proteins designed on the scaffold of the 10th human fibronectin type III domain ((10)Fn3), an extremely stable, single-domain protein with an immunoglobulin-like fold but lacking disulfide bonds. Large libraries of possible target-binding proteins can be constructed on the (10)Fn3 scaffold by diversifying the sequence and length of its surface loops, which are structurally analogous to antibody complementarity-determining regions. Target-binding proteins with high affinity and specificity are selected from (10)Fn3-based libraries using in vitro evolution technologies such as phage display, mRNA display, or yeast-surface display. (10)Fn3-based target-binding proteins have binding properties comparable to those of antibodies, but they are smaller, simpler in architecture, and more user-friendly; as a consequence, these proteins are excellent building blocks for the construction of multidomain, multifunctional chains. The ease of engineering and robust properties of (10)Fn3-based target-binding proteins have been validated by multiple independent academic and industrial groups. In addition to performing well as specific in vitro detection reagents and research tools, (10)Fn3-based binding proteins are being developed as therapeutics, with the most advanced candidate currently in Phase II clinical trials.
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Monomeric proteins based on the synthetic protein (FN3) skeleton are similar in function to antibodies and can be used in combination with the corresponding target; they have smaller molecular weight and clearer structure compared to traditional antibodies, and are more stable in oxidation/reduction conditions, making them a new type of ideal inhibitor.
“Monomeric proteins, based on the synthetic protein (FN3) skeleton, are similar in function to antibodies and can be used in combination with the corresponding target. These proteins have a smaller molecular weight and clearer structure compared to traditional antibodies. Moreover, they are more stable in oxidation/reduction conditions, making them a new type of ideal inhibitor.330,331”
From Recent advances in targeting the “undruggable” proteins: from drug discovery to clinical trials (2023) · cites this paper as [331]
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- Authors
- Koide, S., Koide, A., & Lipovšek, D.
- Affiliations
- University of Chicago (US)
- Bristol-Myers Squibb (United States) (US)
- Venue
- Methods in Enzymology
- Publisher
- Elsevier
- Published
- 2012
- Volume
- 503
- Pages
- 135-156
- ISSN
- 0076-6879 (ISSN-L)
- OpenAlex
- W783708
- Cited by
- 84
- References
- 41
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 4 · 2023: 4 · 2022: 2 · 2021: 5 · 2020: 7 · 2019: 5 · 2018: 9 · 2017: 6 · 2016: 9 · 2015: 8 · 2014: 8 · 2013: 6 · 2012: 6
- FWCI
- 30.07
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Methods in enzymology on CD-ROM/Methods in enzymology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of General Medical Sciences (R01GM090324, R01GM072688, U54GM087519)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Keywords
- fibronectin type III domain, single-domain proteins, immunoglobulin-like fold, phage display, mRNA display, yeast surface display, in vitro evolution, hypervariable region, therapeutic proteins, antibody mimics
- MeSH terms
- Antibodies, Carrier Proteins, Escherichia coli, Fibronectins, Humans, Plasmids, Recombinant Fusion Proteins, RNA, Messenger, Substrate Specificity, Yeasts, Protein Engineering, Drug Delivery Systems, Protein Folding, Peptide Library, Protein Sorting Signals, Complementarity Determining Regions, Protein Stability
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