Fig. 1
From: Modeling antibody drug conjugate potential using a granzyme B antibody fusion protein
GrBmut-TRA construct design and physical characterization. A Schematic representation of DNA vectors required for expressing Pro-GrBmut-TRA as a human IgG1 antibody in Expi293 cells. Following affinity chromatography purification, EK cleavage was required to generate a catalytically active GrBmut-TRA molecule. B SDS-PAGE and Coomassie Blue staining of purified TRA and GrBmut-TRA under reducing and non-reducing conditions. Arrow insets indicate HC and LC bands for each molecule. C Western blot analysis of TRA, Pro-GrBmut-TRA, and GrBmut-TRA using reagents specific to human IgG and GrB. Abbreviations used: EK, enterokinase; GrB, granzyme B; rGrB, recombinant GrB; GrBmut-TRA, catalytically active AFP; HC, heavy chain; LC, light chain; NR, non-reduced; Pro-GrBmut-TRA, catalytically inert molecule; R, reduced; TRA, trastuzumab