title: [Expression and purification of T7 bacteriophage capsid protein P11 and preparation of monoclonal antibody against P11].
abstract: OBJECTIVE
  To express capsid tail protein P11 of T7 bacteriophage and produce mouse monoclonal antibody(mAb) against the protein.
  
  
  METHODS
  P11 protein was cloned and recombinant P11 protein was expressed as a fusion protein with an N-terminal 6-His tag. The purified protein was used to immunize BALB/c mouse. The specificity of mAb was analyzed by ELISA and Western blot.
  
  
  RESULTS
  P11 protein was successfully expressed and purified. SDS-PAGE analysis showed that the molecular weight of the expressed protein was approximately 27 kd. One hybridoma cell (2G11) secreting mAb against P11 was developed. The isotype of the mAb was IgG(2b). ELISA detection showed that titers of mAb was 1:8. 1 x 10(5) in ascites.Western blot analysis proved mAb obtained could react specifically to the recombinant p11 protein.
  
  
  CONCLUSIONS
  Recombinant P11 protein and mAbs were successfully prepared.
authors.affiliation: College of Veterinary Medicine, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, China. cl198211@yahoo.com.cn
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Yun-ying
authors.initials: YY
authors.lastname: Gao
authors.forename: Hua-mao
authors.initials: HM
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Xu
authors.forename: Su-xia
authors.initials: SX
authors.lastname: Liu
authors.forename: Zong-hai
authors.initials: ZH
authors.lastname: Li
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D017123:Bacteriophage T7 / Q000737:chemistry* / Q000235:genetics; D036022:Capsid Proteins / Q000276:immunology*; D002459:Cell Fusion; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D014764:Viral Proteins / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D007074:Immunoglobulin G; D014764:Viral Proteins