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Fig. 5 | BMC Biotechnology

Fig. 5

From: Ex vivo evolution of human antibodies by CRISPR-X: from a naive B cell repertoire to affinity matured antibodies

Fig. 5

Characterization of evolved antibodies against HLA-A2. a ELISA dose-response curves of R2+ mutated mAbs C4.4 and C4.18 compared to A2Ab. Statistical significance was determined using a one-way ANOVA test followed by a Tukey’s multiple comparison post-test (n = 3, bars indicate standard deviations) (****: p < 0.0001; ***:p = 0,003). b The affinity of C4.18 was measured by surface plasmon resonance by flowing various concentrations of pp65-A2 complex over CM5 chip-bound C4.18. c Top panel: staining of 721.221 cells which either express HLA-A2 (721.221(A2)) or do not express it (721.221) by A2Ab, C4.4 and C4.18 at 20 μg/mL. MFI are indicated. Lower panel: dose response staining of A2Ab, C4.4 and C4.18 against BLCL HEN expressing HLA-A2. MFI obtained with various concentrations of C4.18 and C4.4 are indicated. d The specificity of mutated R2+ mAbs was assessed in a Luminex single antigen bead assay. Results are shown in terms of interval MFI. Positivity threshold was set at 1000

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