Fig. 3: Designed 5HCS CTLA-4 binder.
From: Design of high-affinity binders to immune modulating receptors for cancer immunotherapy
a Left: Model of 5HCS_CTLA4_1 (cartoon) binding to CTLA-4 (PDB ID:1l85) colored by Shannon entropy from site saturation mutagenesis results. Right: Log enrichments for the 5HCS_CTLA4_1 SSM library selected with 10 nM CTLA-4 at representative positions. The annotated amino acid in each column indicates the residue from the parent sequence. b Biolayer interferometry characterization of 5HCS_CTLA4_1. Biotinylated CTLA-4 was loaded to Streptavidin (SA) tips, and these were incubated with 2.7 nM, 0.9 nM, and 0.3 nM of 5HCS_CTLA4_1 to measure the binding affinity. c Crystal structure of 5HCS_CTLA4_1 in complex with CTLA-4. Color schemes are the same as Fig. 2c. Designed interactions between 5HCS_CTLA4_1 (green) and CTLA-4 (white). d Circular dichroism spectra from 25 °C to 95 °C for 5HCS_CTLA4_1. Color schemes and experimental details are as in Fig. 2b. e Increase of TCR activation induced signal (via NFAT pathway) from engineered CTLA-4 effector cells lines by 5HCS_CTLA4_1 (green), lpilimumab (gold), and 5HCS_CTLA4_1_c6 (blue) is shown. EC50 values were fitted using four-parameter logistic regression by Python scripts. Source data (b, d, e) are provided in the Source Data file.
