Removal of oral Prevotella intermedia Endotoxin by octyl phenyl polyoxyethylene ether extraction method / 中华口腔医学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate an effective purification method for removing endotoxin from Prevotella intermedia.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>The extraction method provides a new choice for endotoxin removal from large volumes of the oral Prevotella intermedia.</p>