title: Development of high-affinity monoclonal antibody using CD44 overexpressed cells as a candidate for targeted immunotherapy and diagnosis of acute myeloid leukemia.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Amanzadeh
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Heidarnejad
authors.affiliation: Immunology Department, Hybridoma Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Meghdad
authors.initials: M
authors.lastname: Abdollahpour-Alitappeh
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Molla-Kazemiha
authors.affiliation: Department of Mycobacteriology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Shamsi
authors.initials: S
authors.lastname: Yari
authors.affiliation: Department of Mycobacteriology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Hadizadeh-Tasbiti
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Habibi-Anbouhi
authors.affiliation: Immunology Department, Hybridoma Laboratory, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Abolhassani
authors.affiliation: National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Shokrgozar
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D002999:Clone Cells; D005260:Female; D015870:Gene Expression; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000276:immunology*; D006825:Hybridomas / Q000166:cytology / Q000276:immunology; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000302:isolation & purification; D007167:Immunotherapy / Q000379:methods; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis* / Q000276:immunology / Q000473:pathology / Q000628:therapy; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; C497483:CD44 protein, human; D018960:Hyaluronan Receptors; D007074:Immunoglobulin G
keywords: CD44 monoclonal antibody; ascitic fluid; biomarker; high-affinity; hybridoma