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| Main Authors: | , , , , , |
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| Format: | Artículo Open Access |
| Published: |
Wiley
2026
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| Subjects: | |
| Online Access: | https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/bit.70221 |
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Table of Contents:
- Engineered B7‐H3 Binding in Modular Gp2 Miniproteins Abbigael Harthorn Hannah K. Windsor Zachary Schmitz Nathaniel Cheung Ebube Agwaramgbo Benjamin J. Hackel Biotechnology and Bioengineering ABSTRACT B7‐H3, an important immune checkpoint modulator of T‐cell function, is a tumor vasculature biomarker and is overexpressed in a variety of cancers. Its expression is associated with tumor growth, metastasis, and poor clinical prognosis, which makes B7‐H3 an appealing target for diagnostics and therapeutics. High‐affinity, specific, modular ligands are needed to achieve the various modes of molecular targeting strategies. A designed combinatorial library of the small (45 amino acid) Gp2 scaffold was sorted for binders to B7‐H3 via yeast surface display with magnetic and flow cytometric cell sorting. Select variants were sequenced, characterized for binding affinity and specificity to B7‐H3, and assessed for modularity in a protein‐protein fusion. Protein ligands achieved single‐digit nanomolar affinities and retained binding affinity upon conjugation, via a glycine‐rich linker, to an enzyme. Directed evolution resulted in a potent, 0.7 nM binder with increased stability as assessed through the apparent midpoint of denaturation of 67°C. The engineered ligands provide small, modular, high‐affinity B7‐H3 binders for molecularly targeted therapeutics and diagnostics. 10.1002/bit.70221 http://creativecommons.org/licenses/by/4.0/