title: Production and characterization of murine monoclonal antibody against synthetic peptide of CD34.
abstract: BACKGROUND
  The treatment of hematologic malignancies and immunodeficiency diseases are offered by hematopoietic stem cells (HSCs) as a unique self-renewal and differentiation source which most commonly is selected by CD34 surface marker for HSC.
  
  
  OBJECTIVE
  The purpose of this study was to develop and characterize monoclonal antibody against CD34 antigen for detection of hematopoietic stem cells.
  
  
  METHODS
  Balb/c mice were immunized with two synthetic peptides of CD34 and Spleen cells were fused with SP2/0.Fused cells were grown in hypoxanthine, aminopterine and thymidine (HAT) selective medium and cloned by limiting dilution. Large scale of monoclonal antibodies was produced by mouse ascites production of mAb (in vivo) method. Monoclonal antibody was purified by chromatography. Then reactivity of these antibodies was evaluated in different immunological assays including ELISA, immunofluorescence (IF), western blot (WB) and flowcytometry.
  
  
  RESULTS
  In this study, between five positive clone wells, two clones were chosen for limiting dilution. Limiting dilution product was one monoclone (3-D5 monoclone) with absorbance about 2. Isotype of this mAb was identified as IgG1 class with Kappa (κ) light chain.
  
  
  CONCLUSIONS
  This antibody is highly specific and functional in biomedical applications such as ELISA, flowcytometry, immunofluorescence, and western blot assays.
authors.affiliation: Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Leili Aghebati
authors.initials: LA
authors.lastname: Maleki
authors.forename: Jafar
authors.initials: J
authors.lastname: Majidi
authors.forename: Behzad
authors.initials: B
authors.lastname: Baradaran
authors.forename: Jalal
authors.initials: J
authors.lastname: Abdolalizadeh
authors.forename: Aliakbar Movassaghpour
authors.initials: AM
authors.lastname: Akbari
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D000918:Antibody Specificity; D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002459:Cell Fusion; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000302:isolation & purification*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018952:Antigens, CD34; D015415:Biomarkers; D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D010455:Peptides
keywords: CD34; Monoclonal antibody; synthetic peptide