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The vector of anti-CD30 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD30. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD30 antibody linked to CD3ζ signaling domains. And the vector product was designed for the treatment of CD30 positive cancers.
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CAR Construction : HRS3 scFv-41BB-CD28ζ
Fig.1 Engineered human T-cells express the CAR HRS3. Human peripheral T-cells were isolated from the blood of a healthy donor and retrovirally engineered by transduction of vector HRS3. After 12 hrs cells were tested for HRS3 CAR expression by two colour FACS according to FACS using a PE-labelled polyclonal goat anti-human IgG Fc antibody and the FITC-labelled murine anti-human CD3 monoclonal antibody UCHT1 . |
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CAR Construction : HRS3 scFv-41BB-CD28ζ
Fig.2 T-cells with anti-CD30 CAR HRS3 preferentially accumulate during in vitro culture. Peripheral blood lymphocytes were engrafted with the respective CARs. After transduction cells were cultured in Xvivo15 medium supplemented with 10% FCS and IL-2 (500 U/ml). Medium was replaced once a week, IL-2 was added every 3rd day. CD3+ T-cells were recorded for CAR expression utilizing FITC-conjugated anti-mouse and PE-conjugated anti-human lgG1 Fc antibodies at the indicated time after start of culture. |
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