Strategic design to create HER2-targeting proteins with target-binding peptides immobilized on a fibronectin type III domain scaffold
| Autor: | Yuji Ito, Tetsuya Kadonosono, Shinichi Sato, Maika Kitazawa, Takahiro Kuchimaru, Tadashi Shiozawa, Masumi Taki, Yumi Ota, Shinae Kizaka-Kondoh, Hiroyuki Nakamura, Wanaporn Yimchuen |
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| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
0303 health sciences Scaffold medicine.diagnostic_test Chemistry General Chemical Engineering Proteolysis Peptide General Chemistry Fibronectin type III domain Affinity maturation Dissociation constant 03 medical and health sciences 0302 clinical medicine Biochemistry In vivo 030220 oncology & carcinogenesis Cancer cell medicine 030304 developmental biology |
| Zdroj: | RSC Advances. 10:15154-15162 |
| ISSN: | 2046-2069 |
| Popis: | 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 (KD) 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. |
| Databáze: | OpenAIRE |
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