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. Author manuscript; available in PMC: 2023 Apr 20.
Published in final edited form as: Sci Transl Med. 2022 May 18;14(645):eabn0402. doi: 10.1126/scitranslmed.abn0402

Fig. 1. Deriving a second-generation CDP library with improved surface folding characteristics.

Fig. 1.

(A) Illustration of mammalian surface display principle. If the CDP is not tagged (vector SDGF), tagged (e.g., biotin) target along with a fluorescent co-stain (e.g., streptavidin) stains cells to enrich for target-binding CDPs of interest. When the CDP itself is tagged (e.g., 6xHis, vector SDPR), the fluorescent co-stain (e.g., anti-6xHis) will detect intact CDPs on the surface of the cell. (B and C) High surface folding scores (B) from pooled analysis of library NCL1 permitted homology-based selection from a large CDP library (C) to create a second-generation CDP library, NCL2. (D) Flow-based comparison of SDPR-cloned 6xHis-tagged NCL1 and NCL2 with or without trypsin (5 or 20 μg/mL, 5 mins) followed by DTT treatment (5 mins) and subsequent anti-6xHis antibody staining.