title: [Generation and identification of phage engineering antibodies library against hamster prion protein].
abstract: OBJECTIVE
  Generation and Identification of Phage Engineering Antibodies Library against Hamster Prion Protein.
  
  
  METHODS
  Fab antibodies were identified and confirmed. BALB/c mice were immuned with PrP proteins. After the third immunization, the total RNA was extracted from the mice spleens. The genes of heavy Fd Fragments and light chain of antibodies amplified by a series of specific primers of human IgG Fab fragment were cloned into phagemid vector pComb3.
  
  
  RESULTS
  The combinatorial Fab library were constructed successfully and the cloning efficiencies both of light chain and Fd fragments were about near 10(6). The Fab library were panned by four cycles and screened with purified haPrP23-231 antigen on microtiter plates. 12 mAbs were isolated after four cycles of panning, five of which were sequenced and resulted sequence data were analyzed by alignment with GenBank immunoglobulin genes. Two strain of new heavy and light chain genes of Fab antibodies were identified and confirmed.
  
  
  CONCLUSIONS
  The research in this article will provide foundation for study of diagnosis and therapy of prion.
authors.affiliation: State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
authors.forename: Yin-Xia
authors.initials: YX
authors.lastname: Huang
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Chen-Fang
authors.initials: CF
authors.lastname: Dong
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.forename: Chen
authors.initials: C
authors.lastname: Gao
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Wang
authors.forename: Lu
authors.initials: L
authors.lastname: Han
authors.forename: Wei
authors.initials: W
authors.lastname: Zhou
authors.forename: Bao-Yun
authors.initials: BY
authors.lastname: Zhang
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Jiang
authors.forename: Mi-Fang
authors.initials: MF
authors.lastname: Liang
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015153:Blotting, Western; D006224:Cricetinae; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007114:Immunization; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D011328:Prions / Q000235:genetics* / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D011328:Prions