Process for the preparation of genetically modified t cells expressing the chimeric antigen receptor

Improve the gene transfer efficiency in CAR therapy utilizing transposon method. By (2) a transposon method; (1) after stimulation with anti-CD3 antibody and anti-CD28 antibodies cell populations comprising T cells, the step of providing a non-proliferative cells obtained by performing the process of loss of proliferative capacity step obtain genetically modified T cells targeted antigen-specific chimeric antigen receptor gene has been introduced; (3) the genetically modified T cells obtained in non-proliferating cells comprising the steps of prepared (2) were mixed in step (1) step cocultured with stimulated with anti-CD3 antibody and anti-CD28 antibody; (4) a step of recovering the cells after culturing, process for the preparation of genetically modified T cells expressing the chimeric antigen receptor. Further, after stimulation with anti-CD3 antibody and anti-CD28 antibody population of cells comprising (i) T cells, obtained by performing processing to loss of culture and ability to grow in the presence of viral peptide antigens, viral peptide antigens by (ii) the transposon method, the step to obtain a genetically modified T cells targeted antigen-specific chimeric antigen receptor gene has been introduced; steps of providing a non-proliferating cells that retain prepared in (iii) step (i) non genetically modified T cells obtained in proliferating cells and step (ii) are mixed, step co-culturing; (iv) comprises the step of collecting the cells after cultivation, the genetically modified T cells expressing the chimeric antigen receptor preparation method is provided.