A Non-immunogenic Bivalent d-Protein Potently Inhibits Retinal Vascularization and Tumor Growth

PS Marinec, KE Landgraf, M Uppalapati… - ACS Chemical …, 2021 - ACS Publications
PS Marinec, KE Landgraf, M Uppalapati, G Chen, D Xie, Q Jiang, Y Zhao, A Petriello…
ACS Chemical Biology, 2021ACS Publications
We report a general approach to engineering multivalent d-proteins with antibody-like
activities in vivo. Mirror-image phage display and structure-guided design were utilized to
create a d-protein that uses receptor mimicry to antagonize vascular endothelial growth
factor A (VEGF-A). Selections against the d-protein form of VEGF-A using phage-displayed
libraries of two different domain scaffolds yielded two proteins that bound distinct receptor
interaction sites on VEGF-A. X-ray crystal structures of the d-protein/VEGF-A complexes …
We report a general approach to engineering multivalent d-proteins with antibody-like activities in vivo. Mirror-image phage display and structure-guided design were utilized to create a d-protein that uses receptor mimicry to antagonize vascular endothelial growth factor A (VEGF-A). Selections against the d-protein form of VEGF-A using phage-displayed libraries of two different domain scaffolds yielded two proteins that bound distinct receptor interaction sites on VEGF-A. X-ray crystal structures of the d-protein/VEGF-A complexes were used to guide affinity maturation and to construct a heterodimeric d-protein VEGF-A antagonist with picomolar activity. The d-protein VEGF-A antagonist prevented vascular leakage in a rabbit eye model of wet age-related macular degeneration and slowed tumor growth in the MC38 syngeneic mouse tumor model with efficacies comparable to those of approved antibody drugs, and in contrast with antibodies, the d-protein was non-immunogenic during treatment and following subcutaneous immunizations.
ACS Publications
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