title: [The expression of human IL-37 in E.coli and preparation and characterization of mouse anti-IL-37 antibody].
abstract: OBJECTIVE
  To construct a prokaryotic expression system for interleukin-37 (IL-37) and prepare its polyclonal antibody.
  
  
  METHODS
  The gene encoding mature interleukin-37 (IL-37m) was amplified by PCR and subcloned into prokaryotic expression vector pET28a. Then the recombinant plasmid pET28a/IL-37m was transformed into E.coli Rosetta and expressed under IPTG induction. The recombinant IL-37m was purified through Ni²⁺;-NT agarose gel column and the purified recombinant IL-37m was used as immunogen to immunize the BALB/c mouse. The titer and specificity of the mouse anti-IL-37 antibody were analyzed by ELISA, Western blotting and immunohistochemical staining, respectively.
  
  
  RESULTS
  The recombinant IL-37 was successfully expressed and purified, and the mouse anit-IL-37 antibody was successfully prepared. ELISA showed that the titer of the antiserum was 1:128 000. Western blot analysis revealed that the antibody reacted with IL-37 specifically. Immunohistochemical staining detection manifested the antibody could recognize the native IL-37.
  
  
  CONCLUSIONS
  The mouse anti-IL-37 antibody with high titer and specificity was successfully prepared.
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Fuqian
authors.initials: F
authors.lastname: Zhao
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Suhui
authors.initials: S
authors.lastname: He
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Lingge
authors.initials: L
authors.lastname: He
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Miaofen
authors.initials: M
authors.lastname: Lin
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Wang
authors.affiliation: Guangdong Key Laboratory for Medical Molecular Diagnostics, Department of Clinical Immunology, Guangdong Medical College, Dongguan 523808, China.
authors.forename: Zhangquan
authors.initials: Z
authors.lastname: Chen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007114:Immunization / Q000379:methods*; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; C508587:IL37 protein, human; D007375:Interleukin-1; D011994:Recombinant Proteins