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Issue 6, 2011
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Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay

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Abstract

Measurement of environmental endotoxin exposures is complicated by variability encountered using current biological assay methods arising in part from lot-to-lot variability of the Limulus-amebocyte lysate (LAL) reagents. Therefore, we investigated the lot-to-lot repeatability of commercially available recombinant Factor C (rFC) kits as an alternative to LAL. Specifically, we compared endotoxin estimates obtained from rFC assay of twenty indoor dust samples, using four different extraction and assay media, to endotoxin estimates previously obtained by Limulus amebocyte lysate (LAL) assay and amounts of 3-hydroxy fatty acids (3-OHFA) in lipopolysaccharide (LPS) using gas-chromatography mass spectroscopy (GC-MS). We found that lot-to-lot variability of the rFC assay kits does not significantly alter endotoxin estimates in house dust samples when performed using three of the four assay media tested and that choice of assay media significantly altered endotoxin estimates obtained by rFC assay of house dust samples. Our findings demonstrate lot-to-lot reproducibility of rFC assay of environmental samples and suggest that use of rFC assay performed with Tris buffer or water as the extraction and assay medium for measurement of endotoxin in dust samples may be a suitable choice for developing a standardized methodology.

Graphical abstract: Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay

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Publication details

The article was received on 26 Jan 2011, accepted on 12 Apr 2011 and first published on 09 May 2011


Article type: Paper
DOI: 10.1039/C1EM10035A
Citation: J. Environ. Monit., 2011,13, 1739-1745
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    Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay

    J. H. McKenzie, K. U. Alwis, J. E. Sordillo, K. S. Kalluri and D. K. Milton, J. Environ. Monit., 2011, 13, 1739
    DOI: 10.1039/C1EM10035A

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