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    Optimisation of circulating biomarkers of cell death for routine clinical use.

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    Authors
    Greystoke, Alastair
    Cummings, Jeffrey
    Ward, Timothy H
    Simpson, Kathryn L
    Renehan, Andrew G
    Butt, Fouziah
    Moore, David
    Gietema, J
    Blackhall, Fiona H
    Ranson, Malcolm R
    Hughes, Anthony
    Dive, Caroline
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    Affiliation
    Department of Medical Oncology, Clinical and Experimental Pharmacology Group, Paterson Institute for Cancer Research, Manchester, UK.
    Issue Date
    2008-05
    
    Metadata
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    Abstract
    BACKGROUND: M30 and M65 enzyme-linked immunosorbent assays detect circulating cytokeratin 18 fragments released during caspase-dependent or total cell death, respectively, and have potential as biomarkers in epithelial cancers. While these assays have been validated, their robustness for routine clinical use is unknown. PATIENTS AND METHODS: M30 and M65 were measured in matched serum and plasma samples from 31 lung cancer patients and 18 controls. RESULTS: Time allowable between sample acquisition and processing is critical for assays in clinical use. A 4-h delay in processing at room temperature increased M30 (P < 0.0001), an effect minimised by incubation on ice. M30 and M65 in serum were resistant to processing variations including delays. Serum and plasma measurements correlated well although M30 but not M65 was lower in serum (P < 0.0005). Less variation between duplicate assays was observed in serum. Prolonged storage (-80 degrees C) led to increased M30 (12%, 6 months; 34%, 1 year). Sample dilution in the supplied assay diluent proved non-linear, whereas dilution in donor serum or porcine plasma restored linearity up to a ratio of 1 : 6. CONCLUSION: We present recommendations that improve the reliability of these assays for clinical use and recommend serum as the preferred matrix with data more resistant to variations in collection.
    Citation
    Optimisation of circulating biomarkers of cell death for routine clinical use. 2008, 19 (5):990-5 Ann. Oncol.
    Journal
    Annals of Oncology
    URI
    http://hdl.handle.net/10541/66159
    DOI
    10.1093/annonc/mdn014
    PubMed ID
    18304966
    Type
    Article
    Language
    en
    ISSN
    1569-8041
    ae974a485f413a2113503eed53cd6c53
    10.1093/annonc/mdn014
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research
    Clinical and Experimental Pharmacology Group
    Medical Oncology
    Medical Oncology
    School of Cancer and Imaging Sciences

    entitlement

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