Construction of hybridoma cells with IL1RAP as a new marker for leukemia stem cells and detection of its monoclonal antibody / 中国实验血液学杂志

ABSTRACT

This study was aimed to prepare and identify human monoclonal antibody against IL-1 receptor accessory protein (IL1RAP), which is a new identified surface marker for leukemia stem cell (LSC), BALB/c mice were immunized with recombinant hu-IL1RAP and the spleen cells from immunized mice were fused with SP2/0 myeloma cells by conventional hybridoma technique. Positive hybridoma cells were selected and cultured. ELISA and Western blot were used to detect the type, titer and sensitivity of antibody. Peripheral blood mononuclear cells were isolated and used to test the antibody specificity. The results showed that 8 hybridoma cell lines able to stably secrete IL1RAP monoclonal antibodies were obtained and named 3H6E10, 4B6A6, 8G11B5, 9E9F2, 10D8A7, 1C7H7, 1D7G11 and 2D3D3 respectively. These monoclonal antibodies belonging to IgG1/κ type could specifically bind to IL1RAP from peripheral blood mononuclear cells. It is concluded that the hybridoma cell lines with stable secretion of IL1RAP monoclonal antibodies is successfully constructed, thus providing novel ways to effectively clear LSC in the future.