Fig. 2: CDR mutagenesis of HuJovi-1 for TRBC2 specificity.
From: Structure-guided engineering of immunotherapies targeting TRBC1 and TRBC2 in T cell malignancies
a Molecular model of HuJovi-1 (Lys28 mutant; Mut1) in the context of TRBC2, suggesting that this residue can form a hydrogen bond with Lys119 in TRBC2. b ELISA binding of Thr28 mutant (pink) and HuJovi-1 (blue) to TRBC1 (triangles) and TRBC2 (squares). c Molecular model of HuJovi-1 (Lys28/Phe32; Mut2) in the context of TRBC2. d ELISA binding of Lys28/Phe32 Mut2 (green) and HuJovi-1 (blue) to TRBC1 (triangles) and TRBC2 (squares). e Molecular model of HuJovi-1 (Lys28/Phe32/Asn96; Mut3 – KFN) (PDB ID 7AMS). f ELISA binding of KFN (orange) and HuJovi-1 (blue) to TRBC1 (triangles) and TRBC2 (squares). d–f n = 3 technical replicates. Data shown as mean ± SD with non-linear fit, sigmoidal 4PL model. g Generation and interrogation of a phage display library based on HuJovi-1 structural analysis. Top: Amino acid distribution plot of residues in HuJovi-1 CDRs that were randomized to generate the library. Bottom: Frequency distribution of amino acids of 189 unique TRBC2-specific clones that were obtained from the library after selection. h Affinity ELISA showing the 17 highest affinity TRBC2 binding clones obtained after selection. (all antibody aa positions are referring to Kabat numbering scheme12). Source data are provided as a Source Data file.
