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Pyrogen testing of lipid‐based TPN using Mono Mac 6 monocyte cell line and DELFIA
Author(s) -
Moesby L,
Moesby L,
Wind Hansen E
Publication year - 1997
Publication title -
journal of clinical pharmacy and therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.622
H-Index - 73
eISSN - 1365-2710
pISSN - 0269-4727
DOI - 10.1046/j.1365-2710.1997.00110.x
Subject(s) - lipopolysaccharide , immunoassay , chemistry , detection limit , cell culture , secretion , cytokine , pharmacology , microbiology and biotechnology , biochemistry , immunology , medicine , chromatography , biology , antibody , genetics
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 bio??assay showed a correlation between the results of the two assays. DELFIA did not show any cross‐reactivity with the human cytokines TNFα, IL‐1β, IL‐1α, IL‐2, IL‐4, IFNγ, 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.