Construction of hybridoma cells with IL1RAP as a new marker for leukemia stem cells and detection of its monoclonal antibody.
10.7534/j.issn.1009-2137.2013.06.005
- Author:
Kai ZHAO
1
,
2
;
Ling-Ling YIN
1
;
Dong-Mei ZHAO
3
;
Qing-Yun WU
3
;
Chong CHEN
4
;
Bin PAN
4
;
Ling-Yu ZENG
3
;
Yao YAO
3
;
Kai-Lin XU
1
;
Author Information
1. Laboratory of Transplantation Immunology, Xuzhou Medical College
2. Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
3. Laboratory of Transplantation Immunology, Xuzhou Medical College, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
4. Department of Hematology, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
analysis;
Antibody Specificity;
immunology;
Cell Line;
Humans;
Hybridomas;
immunology;
secretion;
Interleukin-1 Receptor Accessory Protein;
immunology;
Mice;
Mice, Inbred BALB C;
Neoplastic Stem Cells;
immunology
- From:
Journal of Experimental Hematology
2013;21(6):1390-1393
- CountryChina
- Language:Chinese
-
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.
