title: [The expression, identification of human HMGB1 B box and preparation of monoclonal antibodies against HMGB1 B box].
abstract: OBJECTIVE
  To express human HMGB1 B box protein and obtain monoclonal antibodies (mAbs) against HMGB1 B box for further study of the function of human HMGB1 protein.
  
  
  METHODS
  pET28-HMGB1 B box plasmid transfected the DH5α, then expressed. And the extracted protein was purified by protein purification system. BALB/c mice were immunized with recombinant human HMGB1 B box protein. Hybridoma cell lines secreting mAb against human HMGB1 B box protein were screened by ELISA and subcloning approach. The characteristics of these mAbs were identified by ELISA and Western blot.
  
  
  RESULTS
  Two hybridoma cell lines (1D2F4E3 and 2D4E3A2) stable secreting specific mAbs were successfully obtained.Western blot exhitited the two mAbs binded specifically to human HMGB1 B box protein. The immunoglobulin (Ig) class of two mAbs belonged to IgG, their titers were 1×10(6);, and the A(450); of mAb1D2F4E3, 2D4E3A2 were 0.324±0.093, 0.296±0.085, respectively.
  
  
  CONCLUSIONS
  Two of high specificity mAbs against human HMGB1 B box protein have been successfully prepared, which laid the foundation for further study of biological function of human HMGB1 protein.
authors.affiliation: Institute of Clinical Laboratory Medicine, Jiangsu University, Zhenjiang 212013, China. szl30@126.com
authors.forename: Zhao-liang
authors.initials: ZL
authors.lastname: Su
authors.forename: Sheng-jun
authors.initials: SJ
authors.lastname: Wang
authors.forename: Cheng-lin
authors.initials: CL
authors.lastname: Zhou
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Chen
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Cai-hua
authors.initials: CH
authors.lastname: Tao
authors.forename: Qi-xiang
authors.initials: QX
authors.lastname: Shao
authors.forename: Hua-xi
authors.initials: HX
authors.lastname: Xu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics; D024243:HMGB1 Protein / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D024243:HMGB1 Protein; D007074:Immunoglobulin G