title: [Preparation and characterization of mouse monoclonal antibody against outer membrane protein P6 of Haemophilus influenzae].
abstract: OBJECTIVE
  To prepare and identify monoclonal antibody against Haemophilus influenzae(Hi) outer membrane protein P6.
  
  
  METHODS
  Recombinant protein P6 as an immunogen was administered intraperitoneally to BALB/c mice. The splenocytes of the mouse were isolated from spleen and hybridized with Sp2/0 myeloma cells. Indirect ELISA was used for screening hybridoma and the number of chromosomes in hybridoma cells was determined by karyotype analysis. The titers and specificity of monoclonal antibodies in their culture supernatant were detected by indirect ELISA. The immunoglobulin class, subclasses and type of the monoclonal antibody were identified with colloidal gold labeled IsoQuick(TM) strips.
  
  
  RESULTS
  Two hybridoma cell lines designated α2G3 and γ2C4 were obtained. Karyotype analysis showed that the chromosome numbers of α2G3 and γ2C4 were 103 and 95, respectively. The highest titers of antibodies in their culture supernatant were 1:256 and 1:512, respectively. Both monoclonal antibodies only reacted with standard or clinical isolated strains of Hi, and they both did not react with other bacteria. A2G3 was IgG2b, and γ2C4 was IgM, both of which were kappa light chains. They could recognize different antigen epitope of protein P6.
  
  
  CONCLUSIONS
  Two hybridoma cell lines producing the monoclonal antibodies against protein P6 of Hi outer membrane are obtained.
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Wen
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Zhou
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Haixia
authors.initials: H
authors.lastname: Qiao
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Xue
authors.initials: X
authors.lastname: Gao
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Jia
authors.affiliation: Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou 075000, China.
authors.forename: Yutuo
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000235:genetics / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D018073:Haemophilus Vaccines / Q000235:genetics / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D007621:Karyotyping; D008297:Male; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D018073:Haemophilus Vaccines; D007074:Immunoglobulin G; D007075:Immunoglobulin M; C069990:P6 outer membrane protein, Haemophilus; D011994:Recombinant Proteins