title: Preparation of monoclonal antibodies against prion proteins with full-length hamster PrP.
abstract: OBJECTIVE
  To prepare the PrP specific monoclonal antibodies (mAbs) that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases.
  
  
  METHODS
  Several BALB/c mice were immunized with recombinant hamster prion protein (HaPrP). Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry.
  
  
  RESULTS
  The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrPSc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross-reactivity with various PrPC from several other mammalian species, including humans and cattles. Immunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPsSc.
  
  
  CONCLUSIONS
  The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases.
authors.affiliation: School of Medicine, Xi'an Jiao-Tong University, Shaanxi, China.
authors.forename: Xin-Li
authors.initials: XL
authors.lastname: Xiao
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Jiang
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Kai
authors.initials: K
authors.lastname: Nie
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Zhou
authors.forename: Yin-Xia
authors.initials: YX
authors.lastname: Huang
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Bao-Yun
authors.initials: BY
authors.lastname: Zhang
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D045744:Cell Line, Tumor; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007114:Immunization; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018621:PrPC Proteins / Q000235:genetics / Q000276:immunology*; D018620:PrPSc Proteins / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011994:Recombinant Proteins