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. 2021 Aug 9;40(19):e107985. doi: 10.15252/embj.2021107985

Figure 1. Phylogenetic relationship of the anti‐SARS‐CoV‐2 Spike VHH antibodies identified and characterized in this study.

Figure 1

  1. The sequences of the isolated VHH antibodies (49 anti‐RBD and 10 anti‐S1ΔRBD nanobodies) were aligned using Clustal Omega (Sievers et al, 2011). The circular phylogram was reconstructed with Dendroscope (Huson & Scornavacca, 2012). Where applicable, sequence classes are indicated. The anti‐RBD nanobodies are colored according to the two RBD epitopes identified in Fig 5 (with epitope 1 in green, epitope 2 in red, and asterisks marking nanobodies included in Fig 5). Anti‐S1ΔRBD nanobodies are colored in magenta. VHH antibodies that did not neutralize SARS‐CoV‐2 are shown in parentheses. See main text and Fig 5 for details.
  2. Vero E6 cells were inoculated with SARS‐CoV‐2, fixed with paraformaldehyde after two days, stained with 30 nM of the indicated Alexa Fluor 488 (AF488)‐labeled anti‐RBD VHH antibodies, and imaged by confocal laser scanning microscopy (CLSM).
  3. Immunofluorescence (IF) staining as in B, but with the indicated AF568‐labeled anti‐S1ΔRBD VHH antibodies.