Characterisation of the 1st SSI purified MBL standard

Inga Laursen, Peter Højrup, Gunnar Houen, Michael Christiansen

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Abstrakt

BACKGROUND: Mannan-binding lectin (MBL) is of importance in innate immunity. MBL-deficiency, the most common immune defect, is significant in several clinical contexts. The request for MBL diagnostic is increasing, hence we developed a high-purity MBL standard assigned with a traceable value. METHODS AND RESULTS: The standard material was produced from human plasma; and the protein concentration determined by amino acid analysis after a preceding desalting. The standard value was assessed by two series of sub-sample analyses from nine vials by the grand mean: 235.7 microg protein/ml (range 191.1-280.3 microg/ml). The loss during desalting was 7% and the protein content 253.4 microg/ml after correction. After SDS/PAGE the MBL content was estimated by densitometric scanning. The MBL band (non-MBL bands being MASPs) comprised 76%. Therefore, the standard was assigned a value of 192.6 microg MBL/ml (range 156.0-229.2 microg/ml). A calibrated time-resolved immuno-flourescence assay was used for stability evaluation of the MBL standard, after transfer from -80 degrees C, showing stability for at least 10 days at 25 degrees C, 14 days at 5 degrees C, and 16 weeks at -20 degrees C. CONCLUSION: The 1st SSI purified MBL standard has been produced, and assigned the value 192.6 microg MBL/ml, traceable to an accurate realisation of the unit.
OriginalsprogEngelsk
TidsskriftClinica Chimica Acta
Vol/bind395
Udgave nummer1-2
Sider (fra-til)159-161
Antal sider2
ISSN0009-8981
DOI
StatusUdgivet - 2008

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