Cell immunotherapy is one of the most promising, fastest growing therapeutic treatments. Chimeric antigen receptor (CAR) is a synthetic molecular construct, which is inserted in T cells by using different methods of gene transfer. That allows lymphocytes T to bind antigens and other receptors and destroy select cells, which makes them excellent for treatment of B-cell leukemias and lymphomas. There has been a rapid development of chimeric antigen receptors that target other diseases. Main goal of the master's thesis is use of electrogenic transfer for expression of CD19 specific CAR molecules in lymphocytes T and their proliferation and characterization. We propagated a plasmid carrying a GFP gene, which we used for optimizing the conditions of electroporation, as well as a set of plasmids carrying a CAR gene and gene for the enzyme transposase. We characterized the cells by using a fluorescent microscope and flow cytometry. Optimal transfection condition for lymphocytes T was as follows – one pulse with a duration of 15 ms at 2400 V. Expression of CAR was proven with flow cytometry and a functional ELISA test, that was used for measuring the concentration of secreted interleukin 2 following cell activation.
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