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. 2017 Apr 10;8:420. doi: 10.3389/fimmu.2017.00420

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Copyright © 2017 Deschaght, Vintém, Logghe, Conde, Felix, Mensink, Gonçalves, Audiens, Bruynooghe, Figueiredo, Ramos, Tanghe, Teixeira, Van de Ven, Stortelers and Dombrecht.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Alignment of human RON (hRON) nanobodies (see also Table S2 in Supplementary Material). The 28 randomly selected candidate hRON-binding nanobodies are identified by the acronym “NGS” followed by a five digit number. The three sequences marked by an asterisk (NGS00003, NGS00020, and NGS00027) are the non-binding sequences from the randomly selected panel of 28. The 35 nanobodies discovered in the conventional screening campaign and predicted by the next-generation sequencing (NGS) analysis are identified by a one or two digit number, followed by a letter, followed by a two digit number. Numbering of alignment positions was done according to the IMGT V-DOMAIN system (26). CDR regions are highlighted in gray. Dots represent residues identical to the top sequence. Dashes represent gaps introduced by the alignment.