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Image Search Results
Journal: Cellular and Molecular Life Sciences: CMLS
Article Title: A new prodrug form of Affibody molecules (pro-Affibody) is selectively activated by cancer-associated proteases
doi: 10.1007/s00018-014-1751-8
Figure Lengend Snippet: Staphylococcal surface display for characterization of the pro-Affibody. a Schematic illustration of the display vector with relevant features indicated. P Lip S. hyicus promoter region; S secretion signal, PP propeptide, Z Affibody molecule, ABP albumin binding protein, used as surface expression reporter tag, XM region for cell wall anchoring. b Schematic picture showing the staphylococcal surface display technology when displaying a monomeric Affibody molecule. Bacterial cells are incubated with fluorescently labeled target and HSA, labeled with a different fluorophore for monitoring of surface expression levels, and then analyzed with flow cytometry. c Schematic picture showing the staphylococcal surface display of a pro-Affibody and hypothetic dot plot from flow cytometry. Bacteria displaying the non-activated pro-Affibody exhibit the signal for surface expression only (1), whereas bacteria displaying pro-Affibody that has been activated by recombinant protease exhibit signals for surface expression as well as target binding (2). d Representative density plot of the non-activated pro-Affibody displayed on S. carnosus, incubated with 20 nM HER2 and analyzed in the flow cytometer, showing the HER2-binding signal on the y-axis and the surface expression level on the x-axis. e Same as in d, but for the pro-Affibody that has been activated with MMP-1. Experiments were performed in duplicates on different days using freshly prepared samples and reagents
Article Snippet:
Techniques: Plasmid Preparation, Binding Assay, Expressing, Incubation, Labeling, Flow Cytometry, Bacteria, Recombinant