Strategic design to create HER2-targeting proteins with target-binding peptides immobilized on a fibronectin type III domain scaffold

Tumor-binding peptides such as human epidermal growth factor receptor 2 (HER2)-binding peptides are attractive therapeutic and diagnostic options for cancer. However, the HER2-binding peptides (HBPs) developed thus far are susceptible to proteolysis and lose their affinity to HER2 in vivo . In this...

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Bibliographic Details
Published in:RSC advances 2020-04, Vol.1 (26), p.15154-15162
Main Authors: Yimchuen, Wanaporn, Kadonosono, Tetsuya, Ota, Yumi, Sato, Shinichi, Kitazawa, Maika, Shiozawa, Tadashi, Kuchimaru, Takahiro, Taki, Masumi, Ito, Yuji, Nakamura, Hiroyuki, Kizaka-Kondoh, Shinae
Format: Article
Language:English
Subjects:
Affinity
Binding
Cancer
Chemistry
Diagnostic systems
Domains
Fibronectin
Growth factors
In vivo methods and tests
Peptides
Phages
Proteins
Scaffolds
Tumors
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Summary:Tumor-binding peptides such as human epidermal growth factor receptor 2 (HER2)-binding peptides are attractive therapeutic and diagnostic options for cancer. However, the HER2-binding peptides (HBPs) developed thus far are susceptible to proteolysis and lose their affinity to HER2 in vivo . In this report, a method to create a HER2-binding fluctuation-regulated affinity protein (HBP-FLAP) consisting of a fibronectin type III domain (FN3) scaffold with a structurally immobilized HBP is presented. HBPs were selected by phage-library screening and grafted onto FN3 to create FN3-HBPs, and the HBP-FLAP with the highest affinity (HBP sequence: YCAHNM) was identified after affinity maturation of the grafted HBP. HBP-FLAP containing the YCAHNM peptide showed increased proteolysis-resistance, binding to HER2 with a dissociation constant ( K D ) of 58 nM in ELISA and 287 nM in biolayer interferometry and specifically detects HER2-expressing cancer cells. In addition, HBP-FLAP clearly delineated HER2-expressing tumors with a half-life of 6 h after intravenous injection into tumor-bearing mice. FN3-based FLAP is an excellent platform for developing target-binding small proteins for clinical applications. A HER2-binding protein, HBP-FLAP, developed by peptide immobilization specifically binds to HER2 and has improved resistance to proteases.
ISSN:2046-2069
2046-2069
DOI:10.1039/d0ra00427h