title: [Preparation and preliminary application of monoclonal antibody against recombinant human erythropoietin].
abstract: OBJECTIVE
  To prepare monoclonal antibody (mAb) against erythropoietin(EPO), characterize its biological properties and use it to purify the rhEPO from transgenic goat milk.
  
  
  METHODS
  A crude rhEPO product was used as the antigen to immunize BALB/c mice for preparing mAbs against rhEPO. The mAbs were characterized by Western blot and indirect ELISA. Purified mAb 2E6 was coupled with pre-activated Sepharose 4B to prepare the immunoaffinity chromatography column for purifying the rhEPO from transgenic goat milk.
  
  
  RESULTS
  Two hybridoma cell lines(1E7 and 2E6)were obtained. mAbs 1E7 and 2E6 were shown to be IgG1 and IgG2b respectively, and their light chains were both kappa. Western blot analysis confirmed that the two mAbs could bind to rhEPO. The immunoaffinity chromatography column could adsorb 70% of rhEPO in purifying the rhEPO from transgenic goat milk.
  
  
  CONCLUSIONS
  Two hybridoma cell lines secreting anti-rhEPO mAbs were successfully established. The mAb-immunoaffinity chromatography column could be used to purify the rhEPO from transgenic goat milk.
authors.affiliation: College of Life Science and Technology, Tongji University, Shanghai 200092, China. yanfangw976@sohu.com
authors.forename: Yan-fang
authors.initials: YF
authors.lastname: Wang
authors.forename: Jin
authors.initials: J
authors.lastname: Peng
authors.forename: Yin-feng
authors.initials: YF
authors.lastname: Cai
authors.forename: Ya-ming
authors.initials: YM
authors.lastname: Ni
authors.forename: Jian-yang
authors.initials: JY
authors.lastname: Zhao
authors.forename: Guo-xiang
authors.initials: GX
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004921:Erythropoietin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006041:Goats; D006825:Hybridomas / Q000276:immunology*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins; D004921:Erythropoietin