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The vector of anti-ALPP chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Human ALPP. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-ALPP antibody linked to 4-1BB and CD3ζ signaling domains. And the vector product was designed for the treatment of Renal cell carcinoma.
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Fig.1 Binding of MS17-57 to lysates of MKN45, BGC823 and GES-1 cell lines in indirect ELISA. MKN45, BGC823, SGC7901, and MKN28 cells were used in experiments, but MKN45 and BGC823 cell lines were randomly selected as examples in this figure. MS17-57 expressed strong binding signals to MKN45 cells and moderate binding signals to BGC823 cells and GES-1 cells. Cell lysates were coated with 1.0 µg/mL PBS onto Immulon-II HB 96-well ELISA plates (100 µL/well). The protein concentrations of these cell lysates were balanced, but not for the binding targets (Ags) that could be a big variation. Irrelevant mAb was used as an isotype control. Li, Ming, et al. "Generation of monoclonal antibody MS17-57 targeting secreted alkaline phosphatase ectopically expressed on the surface of gastrointestinal cancer cells." PloS one 8.10 (2013): e77398. Distributed under Open Access license CC BY 4.0, without modification. |
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Fig.2 Cells from fresh surgical tissues stained with MS17-57 and isotype control mAb. Immunofluorescence cell staining with MS17-57 revealed significantly stronger staining of GI tumor tissues than that of normal (adjacent noncancerous) control tissues (P<0.03 overall). (Data from patient 3 were omitted for analyses because the tissue had not been properly prepared.). Li, Ming, et al. "Generation of monoclonal antibody MS17-57 targeting secreted alkaline phosphatase ectopically expressed on the surface of gastrointestinal cancer cells." PloS one 8.10 (2013): e77398. Distributed under Open Access license CC BY 4.0, without modification. |
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Fig.3 ICC staining for MS17-57 binding to MKN45, BGC823, and GES-1 cells on cytospin slides. Two ICC assays were performed; photomicrographs from one are shown at 40x and images from the other at 100x). MS17-57 bound to all three types of cells. The binding target (marker) was located on the cell surface. Images of blank and negative (isotype) controls were also obtained but are not shown here. Li, Ming, et al. "Generation of monoclonal antibody MS17-57 targeting secreted alkaline phosphatase ectopically expressed on the surface of gastrointestinal cancer cells." PloS one 8.10 (2013): e77398. Distributed under Open Access license CC BY 4.0, without modification. |
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Fig.4 MS17-57 inhibits BGC823 cell migration over 3 days. Dose-dependent MS17-57 versus isotype mAb (irrelevant mAb) and medium controls inhibit BGC823 cell migration in a colorimetric cell migration assay. Li, Ming, et al. "Generation of monoclonal antibody MS17-57 targeting secreted alkaline phosphatase ectopically expressed on the surface of gastrointestinal cancer cells." PloS one 8.10 (2013): e77398. Distributed under Open Access license CC BY 4.0, without modification. |
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