title: Development and characterization of monoclonal antibodies against human CD20 in Balb/c mice.
abstract: BACKGROUND
  Targeting CD20 antigen on B-lymphocytes provides good opportunity for management of the target cells in patients with B-cell malignancies. By the advent of hybridoma technology, monoclonal antibodies applications exert extensive changes in medical fields such as diagnosis, treatment and purification.
  
  
  OBJECTIVE
  The prim aim of this study was to produce monoclonal antibody against CD20 for exploitation in diagnosis.
  
  
  METHODS
  In this study, Balb/c mice were immunized with two peptides from extracellular domain of CD20. Poly Ethylene Glycol (PEG) fused spleen cells of the most immune mouse with SP2/0 (myeloma cells). Supernatant of hybridoma cells were screened for detection of antibody by ELISA. The desired clones were selected for limiting dilution (L.D). Afterward, specificity and cross reactivity of these antibodies were determined by immunological assay such as ELISA and western blot analysis (WB) and Immunofluorescence. Large scale of monoclonal antibodies was produced by ascetic fluid method. Monoclonal antibody was purified by chromatography then confirmed by SDS-PAGE.
  
  
  RESULTS
  In this study, between five positive clone wells, 3 clones were chosen for limiting dilution. Limiting dilution product was one monoclone with absorbance about 2.
  
  
  CONCLUSIONS
  These results indicate that such monoclonal antibodies against CD20 can be used in diagnosis of CD20 in the cells surface.
authors.affiliation: Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Koushan Sineh
authors.initials: KS
authors.lastname: Sepehr
authors.forename: Behzad
authors.initials: B
authors.lastname: Baradaran
authors.forename: Jafar
authors.initials: J
authors.lastname: Majidi
authors.forename: Jalal
authors.initials: J
authors.lastname: Abdolalizadeh
authors.forename: Leili
authors.initials: L
authors.lastname: Aghebati
authors.forename: Fatemeh Zare
authors.initials: FZ
authors.lastname: Shahneh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D018951:Antigens, CD20 / Q000032:analysis* / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology; D011092:Polyethylene Glycols / Q000737:chemistry; D017434:Protein Structure, Tertiary; D012680:Sensitivity and Specificity; D013154:Spleen / Q000166:cytology / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D010455:Peptides; D011092:Polyethylene Glycols