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. Author manuscript; available in PMC: 2023 Sep 5.
Published in final edited form as: ACS Synth Biol. 2021 Feb 3;10(2):379–390. doi: 10.1021/acssynbio.0c00566

Figure 1.

Figure 1.

Recombinant N bait protein, sdAb sequences derived from panning and their preliminary binding characteristics. (A) Coomassie stained SDS-PAGE analysis of IMAC pure recombinant N protein derived from E. coli expression. Molecular weight markers in kDa; 1, lysate pre-IMAC; 2, flow-through; 3-7, alternate peak fractions; N predicted mwt. 46.7 kDa. (B) Predicted amino acid sequences of the two sdAb selected on N highlighting the complementarity determining regions (CDRs). These Nomad derived clones have additional EPKTPKPQPAASGAEFAAA sequence after FR4 encoding the long hinge, Sfi I and Not I restriction sites. (C) Titration of sdAb-Gluc fusion proteins over constant amount of purified NP protein to determine the EC50 +/− SD shown in the legend. Data points are the means of two separate experiments with error bars representing +/− SD. (D) Western blot of HEK293T lysates following transient expression of recombinant N or control beta-galactosidase (β-gal) genes in pcDNA 48 h post transfection, probed with sdAb-APEX2 fusions and developed with DAB. Molecular weight markers are in kDa.