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. 2020 Aug 10;12(9):e12739. doi: 10.15252/emmm.202012739

Figure 4. Identification of anti‐PrP Fab71 analogous antibodies in human antibody repertoires.

Figure 4

  • A
    Fab71‐similar HCDR3 sequences in three different NGS datasets (DW: dataset for VH:VL; BB and DK datasets for VH) of human antibody repertoires from healthy donors. Pie chart and stacked bar plot (middle) indicate the occurrence of the identified Fab71 similar HCDR3s: 555 out of 629 identified sequences were only found in one donor, while 74 sequences were shared between donors and occurred in 2 up to six donors. Stacked bar plots (right) report the number of sequences differing to Fab71 HCDR3 by 3, 2 or 1 residues. In total, 13 sequences differed by only one residue. Among them, four sequences were shared between different donors.
  • B
    Sequence alignment of Fab71 VH3–30 with HCDR3 regions from the different databases that differ from Fab71 by ≤ 3 residues (upper panel). Amino acids differing from Fab71 are highlighted by orange boxes. Lower panel: sequence alignment of Fab71 light‐chain VK3–15 with DK_VL2–14 that is naturally paired with DK_VH3‐07.
  • C
    Left: Heat map showing the binding specificities (Z‐scores) of selected Fab71 human analogous antibodies to human recPrP23‐230 compared to the negative controls (BSA and Neu). Right: Heat map representing the reactivity (−logEC50) obtained from dose‐dependent ELISA binding curves of the analogous antibodies to human recPrP23‐230. Red: low reactivity; blue: high reactivity.