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. 2019 Dec 26;9(1):68. doi: 10.3390/jcm9010068

Figure 1.

Figure 1

Design, expression and RIP activity of scFvCD133/rGelonin. (a) Schematic representation of the recombinant scFvCD133/rGelonin fusion construct including the enterokinase (EK) and thrombin cut sites. (b) Coomassie Blue stain of non-cut (NC) and EK cut material with increasing concentration of enterokinase. (c) Coomassie Blue stain of non-cut and thrombin cut (T-cut) material, the material left on resin (LOR) upon His-tag removal (concentrated), and the final cut and purified product (Final). Increasing concentrations of bovine serum albumin (BSA) were used as control. Inserted panel under the BSA bands shows an attempt on cutting the final product with EK upon T-cut. (d) Western blot immunodetection of scFvCD133/rGelonin and rGelonin. The immunoblot is representative of three individual experiments with different toxin concentrations. (e) Ribosome-inactivating protein activity of scFvCD133/rGelonin and rGelonin measured by using the cell free Rabbit reticulocyte lysate assay. Representative single well-based data from three independent experiments.