Target-binding proteins based on the 10th human fibronectin type III domain (¹⁰Fn3).
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.
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois, USA.
Shohei
S
Koide
Akiko
A
Koide
Daša
D
Lipovšek
D000906:Antibodies / Q000737:chemistry; D002352:Carrier Proteins / Q000737:chemistry* / Q000627:therapeutic use; D022801:Complementarity Determining Regions / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry; D005353:Fibronectins / Q000737:chemistry* / Q000627:therapeutic use; D006801:Humans; D019151:Peptide Library; D010957:Plasmids / Q000737:chemistry; D015202:Protein Engineering; D017510:Protein Folding; D021382:Protein Sorting Signals; D055550:Protein Stability; D012333:RNA, Messenger / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000627:therapeutic use; D013379:Substrate Specificity; D015003:Yeasts / Q000737:chemistry
D000906:Antibodies; D002352:Carrier Proteins; D022801:Complementarity Determining Regions; D005353:Fibronectins; D019151:Peptide Library; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C501364:fibronectin type III like peptide, human