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. 2021 Nov 11;11:22098. doi: 10.1038/s41598-021-01603-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Antibody-guided screening for Zif-FLAP candidates. (a) Diagram of the Zif-FLAP candidate library construct. Zif lib, Zif-FLAP candidate peptide library. (b) Amino acid frequency in each library set prior to screening. (c) Isolation of the high sfGFP, low AF647 fraction after flow cytometric analysis. K562/CD25 cells transduced with the library construct were incubated with daclizumab-AF647 and then analyzed by flow cytometry for sfGFP expression and AF647 signals. The quadrant in yellow represents the fraction that was sorted out for sequencing. (d) Ranking of peptides in accordance with the read count after NGS analysis. The five most frequently detected sequences are shown by blue circles. The basiliximab (Bas)-derived sequence is indicated by a red circle. (e) Sequences of the top five peptides with the highest read counts. Residues that had been mutated are shown in bold with residues in black representing those that had been fully randomized and those in red representing the CD25-binding residues identified from daclizumab.