1. A Non-immunogenic Bivalent d-Protein Potently Inhibits Retinal Vascularization and Tumor Growth
- Author
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Kyle E Landgraf, Maruti Uppalapati, Qiyang Jiang, Annalise Petriello, Stephen B. H. Kent, Gang Chen, Paul S Marinec, Daniel Xie, Dana Ault-Riche, Sachdev S. Sidhu, Kurt Deshayes, and Yanlong Zhao
- Subjects
0301 basic medicine ,Models, Molecular ,Vascular Endothelial Growth Factor A ,Phage display ,Protein Conformation ,Drug Evaluation, Preclinical ,Antineoplastic Agents ,Eye ,01 natural sciences ,Biochemistry ,Affinity maturation ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,In vivo ,Peptide Library ,Neoplasms ,Animals ,Humans ,Amino Acid Sequence ,Receptor ,Binding Sites ,biology ,010405 organic chemistry ,Antagonist ,Retinal Vessels ,Retinal ,General Medicine ,0104 chemical sciences ,Cell biology ,Bevacizumab ,Vascular endothelial growth factor A ,030104 developmental biology ,Receptors, Vascular Endothelial Growth Factor ,chemistry ,biology.protein ,Molecular Medicine ,Female ,Rabbits ,Antibody ,Protein Multimerization ,Peptides ,Protein Binding - Abstract
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.
- Published
- 2021