Figure 3
From: Nanobodies targeting ABCC3 for immunotargeted applications in glioblastoma
Evaluation of the targeting potential of NbA42 and NbA213 against ABCC3. (a) NbA42 and NbA213 specifically target ABCC3-expressing cells (A549 WT) over A549ABCC3KO control cell line by flow cytometry. Both cell lines were combined at different proportions. Percentage of targeted cells, relative to the total number of cells, is shown for each proportion (X axis). Theoretical percentages of targeted cells, given by each ratio, have been included (dotted line). (b) Binding affinity of NbA42 and NbA213. Their equilibrium dissociation constant (KD) was calculated by flow cytometry (n = 2). Increasing concentrations of FITC-labeled nanobodies were used to quantify ABCC3 expression in A549 WT and A549ABCC3KO control cell lines. (c) Binding specificity of NbA42 and NbA213. The selective binding to the ABCC3 peptide used for their isolation by biopanning (e.g., ABCC3-A, ABCC3-K and ABCC3-L) was calculated by indirect ELISA. The dark dotted line represents the background signal yielded by the negative control (0.1 M NaHCO3 coating buffer without peptide) (n = 2). (d) Dual-nanobody flow cytometry to evaluate synergies in the detection of ABCC3 in vitro.
