Journal: Journal of cancer therapy

Article Title: Saporin Conjugated Monoclonal Antibody to the Transcobalamin Receptor TCblR/ CD 320 Is Effective in Targeting and Destroying Cancer Cells

doi: 10.4236/jct.2013.46122

Figure Lengend Snippet: TCblR expression profile and concentration of mAb-Saporin required for inhibiting cell proliferation.

Article Snippet: SW48 [SW-48] , ATCC CCL-231 , colorectal adenocarcinoma, epithelial , 35 , 20 , 37 , 18 , 10 , 7 , <0.05 , 1.7 , <0.05.

Techniques: Expressing, Concentration Assay, Binding Assay

Journal: Pharmaceutics

Article Title: The Diphtheria Toxin Translocation Domain Impairs Receptor Selectivity in Cancer Cell-Targeted Protein Nanoparticles

doi: 10.3390/pharmaceutics14122644

Figure Lengend Snippet: Modular organization of T22-empowered proteins. ( A ) Modular disposition and 3D models of T22-GFP-TD-H6 and T22-DITOX-H6. T22-GFP-TD-H6 was engineered to mimic the CXCR4-targeted toxin T22-DITOX-H6 through the replacement of the catalytic domain of the toxin (CD) by the green fluorescent protein (GFP). The translocation domain (TD) was maintained. Both proteins display, from the N to the C terminus, the CXCR4 ligand T22, a flexible linker (L; GGSSRSS), two furin cleavable sites (F; GNRVRRSV) recruited from the original DT, and the hexa-histidine tag (H6), all represented in color code. Furin cleavable sites have proved to be functional in this family of modular constructions . Models in 3D were predicted in AlphaFold2 and represented with ChimeraX. ( B ) The modular disposition of the DT and T22-GFP-H6 are also shown as references. In the center is the T22-GFP-H6 nanoparticle as previously modelled , with each monomer represented in a different color and the stabilizing divalent cations as red spheres (reprinted from with permission from Elsevier). ( C ) Molecular weight and isoelectric point of the three proteins used in this study, predicted by ProtParam (Expasy, http://web.expasy.org/protparam ; accessed on 20 November 2022).

Article Snippet: The performance of T22-GFP-TD-H6 was studied in vitro in CXCR4 + HeLa (ATCC-CCL2) cells and CXCR4 − SW1417 (ATCC-CCL238) cells.

Techniques: Translocation Assay, Functional Assay, Molecular Weight

Journal: Pharmaceutics

Article Title: The Diphtheria Toxin Translocation Domain Impairs Receptor Selectivity in Cancer Cell-Targeted Protein Nanoparticles

doi: 10.3390/pharmaceutics14122644

Figure Lengend Snippet: Cell interactivity and fate of T22-GFP-TD-H6 nanoparticles. ( A ) Time-dependent intracellular accumulation of T22-GFP-H6 and T22-GFP-TD-H6 in HeLa cells. Both proteins (stored in 166 mM NaHCO 3 , 333 mM NaCl, pH 8.0) were incubated at 2000 nM. ( B ) Intracellular accumulation kinetics of T22-GFP-H6 and T22-GFP-TD-H6 with chloroquine (0.5 M) incubated 4 h before protein addition. For comparison purposes, conditions tested were the same as in A. For both ( A , B ), fluorescence values are referred to the background of non-treated cells. Data are shown as mean ± standard error. Significant statistical differences between data pairs (T22-GFP-H6 vs. T22-GFP-TD-H6 at a respective time) are represented by * at p < 0.05. ( C ) CXCR4-specificity of 1000 nM and 2000 nM T22-GFP-H6 or T22-GFP-TD-H6 in HeLa cells, measured by cell internalization after 1 h of exposure. Cells were previously incubated with the CXCR4 antagonist AMD3100 at ten-fold the protein concentration (10 µM or 20 µM). Data are shown as mean ± standard error. Statistical significance between T22-GFP-H6 and T22-GFP-TD-H6 specificity is represented by * p < 0.05. ( D ) Intracellular fluorescence of CXCR4 + HeLa cells and CXCR4 − SW1417 cells exposed to T22-GFP-TD-H6 for 24 h. Fluorescence values are referred to the background of non-treated cells. Data are shown as mean ± standard error. Significant statistical differences between values at each condition and background fluorescence of untreated cells are represented by * at p < 0.05.

Article Snippet: The performance of T22-GFP-TD-H6 was studied in vitro in CXCR4 + HeLa (ATCC-CCL2) cells and CXCR4 − SW1417 (ATCC-CCL238) cells.

Techniques: Incubation, Comparison, Fluorescence, Protein Concentration