Development of high-affinity monoclonal antibody using CD44 overexpressed cells as a candidate for targeted immunotherapy and diagnosis of acute myeloid leukemia. 
CD44s antigens have been suggested as an efficient biomarker for cancer stem cells.  Current study aimed to develop a hybridoma that producing a high affinity murine anti-human CD44 monoclonal antibody for early diagnostic laboratory tests of some cancer.  To make hybridoma against CD44, mice were immunized with MDA-MB-468 cells.  Resulted hybridomas using three culture media were screened by indirect ELISA, then cloned by limiting dilution, and isotype was determined after obtaining ascitic fluid and antibody purification.  We obtained a stable secreting clone, capable of secreting a high-affinity monoclonal antibody against CD44 protein, IgG2a kappa, with the affinity of 5.4 × 10-8 M without cross-reactivity.  We could establish a hybridoma in a native form.  This stable and high-affinity anti-CD44 mAb has a potential for diagnostic procedures and laboratory research.  Thus, it could be exploited as a suitable tool for target-specific diagnosis and even treatment in several cancers.