![Recognition of recombinant HLA-E–peptide complexes by the mAb 4D7. (A) ELISA demonstrating the binding specificity of 4D7 or 3D12 to wells coated with 2 μg/mL of recombinant HLA-E[pHLAG] (blue bar), HLA-E [pHSP60] (light gray bar), or pHLA-G (dark gray bar). Murine IgG1κ isotype (2 μg/mL) served as the control. Absorption was measured at O.D. 650 nm. Error bars represent ± standard deviation (SD). Significance was tested by a 2-way analysis of variance (ANOVA). ∗∗∗∗P < .0001. (B) A ProteOn array showed the affinity of 4D7 at concentrations ranging from 0 to 40 nmol/L to 5 μg of HLA-E[pHLA-G]. Data were analyzed using the equilibrium model. (C-D) Binding model comparison of the variable chain of mAb 4D7 using ClusPro2 web server, depicted in blue and green (PDB ID: 3CDG), represented in shades of gray for the HLA-E alpha chain, and purple for the β2 microtubulin, in the presence (C) and absence (D) of the VMAPRTLFL peptide shown in red. O.D., optical density; W/o, without.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/9/21/10.1182_bloodadvances.2025016276/1/blooda_adv-2025-016276-gr1.jpeg?Expires=1764861798&Signature=GptrWPLM0xZ6uPJjBV63ckkNB31e9ZZuqwP~ipX~189Qx07fw~LfBc6l4TMinrcAZAmMJhFjhsEhsH~E9rUZIJo7LJIoHhmtAakDIa4D6zBSZB1vydPo~-zMtDmlMViv7O5fit5j3JuXDxG7a3j3ek46t9d~hzVMY~APSfnGFg~3p5izuQYCNcg2qG3qS-UqbPkHTAZkohUfz5jV52xy2b1mV2nKfPwyCO6sKKup5JzgY6bX5wzPoKKtEblcxDwPdVWw4VzlD5oum2SX~IHLDVeYgMCxuPekdIhlfsI~LcbAEtFkn7rFnIRnNxA0Q177j-iU3Vk9uW~Ses0dkMQoZA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Recognition of recombinant HLA-E–peptide complexes by the mAb 4D7. (A) ELISA demonstrating the binding specificity of 4D7 or 3D12 to wells coated with 2 μg/mL of recombinant HLA-E[pHLAG] (blue bar), HLA-E [pHSP60] (light gray bar), or pHLA-G (dark gray bar). Murine IgG1κ isotype (2 μg/mL) served as the control. Absorption was measured at O.D. 650 nm. Error bars represent ± standard deviation (SD). Significance was tested by a 2-way analysis of variance (ANOVA). ∗∗∗∗P < .0001. (B) A ProteOn array showed the affinity of 4D7 at concentrations ranging from 0 to 40 nmol/L to 5 μg of HLA-E[pHLA-G]. Data were analyzed using the equilibrium model. (C-D) Binding model comparison of the variable chain of mAb 4D7 using ClusPro2 web server, depicted in blue and green (PDB ID: 3CDG), represented in shades of gray for the HLA-E alpha chain, and purple for the β2 microtubulin, in the presence (C) and absence (D) of the VMAPRTLFL peptide shown in red. O.D., optical density; W/o, without.
