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. 2023 Sep 14;9(9):1835–1845. doi: 10.1021/acscentsci.3c00625

Figure 2.

Figure 2

Development of an anti-XCR1 scFv. (A, B) Design of the anti-XCR1 scFv, which encodes variable heavy (VH) and light (VL) chains from a parent anti-XCR1 IgG (MARX10). The scFv also contains a hydrophilic region (G4S)4 between the VH and VL chains and a sortase recognition tag (LPSTG2H6) that enables protein ligation. (C) Coomassie-visualized SDS-PAGE gel of the recombinant scFv throughout the bacterial expression and purification steps: (1) whole cell lysate; (2) Ni NTA purification; (3) treatment with SUMO protease; and (4) ion-exchange (HiTrap Q HP) chromatography. (D) Linker peptide 1, which included 1a (R = α-bromoacetyl group) and 1b (R = Alexa Fluor 647 (AF647)).