title: [Preparation and characterization of monoclonal antibodies against the hemagglutinin of H9 subtype of avian influenza virus].
abstract: OBJECTIVE
  To prepare monoclonal antibodies (mAb) against the hemagglutinin(HA) of H9 subtype of avian influenza virus (AIV).
  
  
  METHODS
  8 week-old female BALB/c mice were immunized with the inactivated vaccine of H9 subtype of AIV. Splenocytes from the immunized mice were fused with Sp2/0 myeloma cells, and positive hybridoma clones were screened by indirect ELISA and hemagglutination inhibition test (HI). The specificity of the mAb was characterized by ELISA, HI test, indirect immunofluorescence (IF) staining and Western blot.
  
  
  RESULTS
  Three hybridoma cell lines named 2H1, 2A3 and 1C8 against HA of AIV H9 were obtained. The HI titers of 3 mAbs were 1 x 2(8)-1 x 2(13), and the ELISA titers were 1 x 10(-7), 1 x 10(-5) and 5 x 10(-6), respectively. The immunoglobulin subclass of all 3 mAbs was IgG1. Western blot analysis confirmed that mAb 2H1 could recognize HA and reacted to 31 out of 32 isolates of H9 subtype of AIV.
  
  
  CONCLUSIONS
  Three mAbs recognizing HA of H9 subtype of AIV were obtained, which may provide an useful tool for the antigenic analysis, the serological diagnosis, the epidemiological survey and the evaluation of AIV vaccine.
authors.affiliation: Key Laboratory of Animal Lemology, Ministry of Agriculture/College of Animal Science and Veterinary Medicine, Yangzhou University 225009, China.
authors.forename: Yan-bing
authors.initials: YB
authors.lastname: Zeng
authors.forename: Xin-an
authors.initials: XA
authors.lastname: Jiao
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Pan
authors.forename: Jin-lin
authors.initials: JL
authors.lastname: Huang
authors.forename: Ping-hu
authors.initials: PH
authors.lastname: Zhang
authors.forename: Su-hua
authors.initials: SH
authors.lastname: Zhang
authors.forename: Quan-yun
authors.initials: QY
authors.lastname: Sun
authors.forename: Xiu-fan
authors.initials: XF
authors.lastname: Liu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000032:analysis; D009980:Influenza A virus / Q000145:classification / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007074:Immunoglobulin G; C109963:hemagglutinin, avian influenza A virus