title: [Removal of oral Prevotella intermedia Endotoxin by octyl phenyl polyoxyethylene ether extraction method].
abstract: OBJECTIVE
  To investigate an effective purification method for removing endotoxin from Prevotella intermedia.
  
  
  METHODS
  The main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments.
  
  
  RESULTS
  Western blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P < 0.05).
  
  
  CONCLUSIONS
  The extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.
authors: Department of Stomatology, Beijing Chao-Yang Hospital, Capital Medical University, Beijing 100020, China.
authors: Ai-wu
authors: AW
authors: Wang
authors: Yan
authors: Y
authors: Liu
authors: Kong-xin
authors: KX
authors: Hu
authors: Qian
authors: Q
authors: Cheng
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification; D004731:Endotoxins / Q000302:isolation & purification*; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D053582:Interleukin-1alpha / Q000097:blood; D015850:Interleukin-6 / Q000097:blood; D016209:Interleukin-8 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry*; D018713:Prevotella intermedia / Q000737:chemistry* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000097:blood
chemical_list: D001426:Bacterial Proteins; D004731:Endotoxins; D053582:Interleukin-1alpha; D015850:Interleukin-6; D016209:Interleukin-8; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40