Pyrogen testing of lipid-based TPN using Mono Mac 6 monocyte cell line and DELFIA

J Clin Pharm Ther. Oct-Dec 1997;22(5-6):327-33. doi: 10.1111/j.1365-2710.1997.tb00014.x.

Abstract

Objective: Measurement of lipopolysaccharide (LPS) induced interleukin-6 (IL-6) secretion in Mono Mac 6 cells.

Method: Dissociation-enhanced lanthanide fluoro immunoassay (DELFIA), a technique based on time-resolved fluorescence.

Results: A comparison of DELFIA and the B9 bioassay showed a correlation between the results of the two assays. DELFIA did not show any crossreactivity with the human cytokines TNFalpha, IL-1beta, IL-1alpha, IL-2, IL-4, IFNgamma, murine IL-6 or LPS. In aqueous solution the detection limit was 2.5 pg LPS/ml, and in a 25% dilution of a lipid-based total parenteral nutrition (TPN) product it was 25 pg LPS/ml. TPN did not interfere with the DELFIA assay, but the IL-6 secretion from the Mono Mac 6 cells was inhibited at TPN concentrations above 25%.

Conclusion: A combination of Mono Mac 6 cells with DELFIA was found to be a reliable method for detecting LPS, but further validation by independent laboratories is justified.

Publication types

  • Comparative Study

MeSH terms

  • Biological Assay
  • Cell Line
  • Cross Reactions
  • Drug Contamination*
  • Europium
  • Fluoroimmunoassay / methods
  • Humans
  • Indicators and Reagents
  • Interleukin-6 / analysis
  • Lipopolysaccharides / analysis*
  • Monocytes / chemistry*
  • Parenteral Nutrition, Total*
  • Pyrogens / analysis*
  • Quality Control

Substances

  • Indicators and Reagents
  • Interleukin-6
  • Lipopolysaccharides
  • Pyrogens
  • Europium