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Fig. 1 | BMC Biotechnology

Fig. 1

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

Fig. 1

Isolation and characterization of human mAb A2Ab. a Sorting strategy used to isolate HLA-A2-specific B lymphocytes from donor NO. Cells with the following phenotypic characteristics: CD3-, CD19+ (left panel), both PE and APC labeled HLA-A2 tetramers+ (middle panel), HLA-B7 tetramer BV421- (right panel) were isolated and used to produce recombinant antibodies. b A2Ab Ab in Fig. 1b and a control anti- pp65-HLA-A*02:01 human mAb (Ac-anti pp65-A2) were tested by ELISA against the following peptide-MHC recombinant monomers: pp65-HLA-A*02:01 (pp65-A2), MelA-HLA-A*02:01 (MelA-A2) and pUV-HLA-B*0701 (pUV-B7). Statistical significance was determined using a two-way ANOVA test followed by a Tukey’s multiple comparison post-test (n = 3, bars indicate standard deviations) (****: p < 0.0001; *:p = 0,0143; ns: not significant). c) The specificity of A2Ab was assessed in a Luminex single antigen bead assay. Results are shown in terms of interval MFI. Positivity threshold was set at 1000. d The affinity of A2Ab was measured by surface plasmon resonance by flowing various concentrations of pp65-A2 complex over CM5 chip-bound A2Ab

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