9EPA

Crystal structure of DARPin NY_1

Summary for 9EPA

• Entry DOI: 10.2210/pdb9epa/pdb
• Descriptor: DARPin NY_1 (2 entities in total)
• Functional Keywords: darpin targeting mhc molecules in complex with tumor-associated peptide antigens, immune system
• Biological source: synthetic construct
• Total number of polymer chains: 1
• Total formula weight: 18596.22

Authors

Schulte, T.,Sandalova, T.,Walser, M.,Mueller, S.,Venetz, N.,Achour, A. (deposition date: 2024-03-18, release date: 2025-03-26, Last modification date: 2025-12-10)

Primary citation

Venetz-Arenas, N.,Schulte, T.,Muller, S.,Wallden, K.,Fischer, S.,Resink, T.,Kadri, N.,Paladino, M.,Pina, N.,Radom, F.,Villemagne, D.,Bruckmaier, S.,Cornelius, A.,Hospodarsch, T.,Alici, E.,Ljunggren, H.G.,Chambers, B.J.,Han, X.,Sun, R.,Carroni, M.,Levitsky, V.,Sandalova, T.,Walser, M.,Achour, A.
Development of DARPin T cell engagers for specific targeting of tumor-associated HLA/peptide complexes.
Iscience, 28:113926-113926, 2025

PubMed Abstract: The balance between affinity and specificity in T cell receptor (TCR)-dependent targeting of HLA-restricted tumor-associated antigens presents a significant challenge for immunotherapy development. T cell engagers that circumvent these limitations are therefore of particular interest. We established a process to generate bispecific designed ankyrin repeat proteins (DARPins) that simultaneously target HLA-I/peptide complexes and CD3e. These high-affinity T cell engagers elicited CD8 T cell activation against tumor targets with strong peptide specificity, as confirmed by X-scanning mutagenesis and functional killing assays. A cryo-EM structure of the ternary DARPin/HLA-A∗0201/NY-ESO1 complex revealed a rigid, concave DARPin surface spanning the full length of the peptide-binding cleft, contacting both α-helices and the peptide. The present findings reveal promising immuno-oncotherapeutic approaches and demonstrate the feasibility of rapidly developing DARPins with high affinity and specificity for HLA/peptide targets, which can be readily combined with a new generation of anti-CD3e-specific DARPins.

PubMed: 41321628
DOI: 10.1016/j.isci.2025.113926

Experimental method

X-RAY DIFFRACTION (1.76 Å)