Ultrafiltration to remove endotoxins and other cytokine-inducing materials from tissue culture media and parenteral fluids.
The presence of small amounts of endotoxins are often undesirable when investigating cytokines such as interleukin-1 and tumor necrosis factor alpha. Polymyxin B, widely used to block endotoxins, does not block several forms of endotoxins, and at high concentrations, polymyxin B itself stimulates interleukin-1 production. Human peripheral blood mononuclear cells are highly sensitive to endotoxins; they respond with cytokine production to endotoxins at concentrations of 10-50 pg/ml and detect pyrogenic materials nonreactive in the Limulus test. In the present study, ultrafiltration using polysulfone filters was found to remove all interleukin-1- and tumor necrosis factor alpha-inducing substances produced in E. coli cultures. Interleukin-1- and tumor necrosis factor alpha-inducing substances derived from Pseudomonas aeruginosa cultures were also rejected by the filters. Ultrafiltration is therefore a convenient and effective procedure to remove interleukin-1 and tumor necrosis factor alpha-inducing substances from parenteral fluids and solutions that come in contact with blood such as fluids used in hemodialysis. This technique is also applicable for the large-scale production of culture media for mammalian cell expression of recombinant, pyrogen-free proteins intended for use in humans.
Schindler
R
R
Tufts University School of Medicine.
Dinarello
C A
CA
D001685:Biological Factors / Q000302:isolation & purification*; D015152:Blotting, Northern; D003470:Culture Media; D016207:Cytokines; D004731:Endotoxins / Q000302:isolation & purification*; D004926:Escherichia coli; D007375:Interleukin-1 / Q000302:isolation & purification; D015850:Interleukin-6 / Q000302:isolation & purification; D010288:Parenteral Nutrition; D011550:Pseudomonas aeruginosa; D012313:RNA / Q000302:isolation & purification; D014409:Tumor Necrosis Factor-alpha / Q000302:isolation & purification; D014462:Ultrafiltration / Q000379:methods
D001685:Biological Factors; D003470:Culture Media; D016207:Cytokines; D004731:Endotoxins; D007375:Interleukin-1; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D012313:RNA