A Novel Fluorochrome Linked Immunosorbent Assay (FLISA) for the complete analysis of the mannose binding lectin (MBL) complement pathway

In addition to recognizing pathogen‐associated molecular patterns, early components of the MBL‐dependent lectin complement pathway (LCP) also recognize self‐antigens and/or IgM following oxidative stress. Current assays for MBL lack the ability to assess all components of the LCP in a single assay s...

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Bibliographic Details
Published in:The FASEB journal 2007-04, Vol.21 (5), p.A181-A182
Main Authors: Walsh, Mary Christine, Shaffer, Lisa A, Body, Simon C, Shernan, Stanton K, Fox, Amanda A, Collard, Charles D, Taylor, Ronald P, Stahl, Gregory L
Format: Article
Language:English
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Summary:In addition to recognizing pathogen‐associated molecular patterns, early components of the MBL‐dependent lectin complement pathway (LCP) also recognize self‐antigens and/or IgM following oxidative stress. Current assays for MBL lack the ability to assess all components of the LCP in a single assay system. In the present study, we developed a novel, low volume, FLISA that quantitatively assesses the functional status of MBL, MASP‐2 and C3 convertase on mannan‐coated plates from sera/plasma samples. Specificity of functional MBL (in ng/ml) is demonstrated by blockade with anti‐MBL mAb 3F8. Additionally, functional MASP‐2 and C3 convertase activity is demonstrated by C4b (pg) and C3b (pg) deposition, respectively in the same wells and are proportionally correlated to functional MBL levels. Furthermore, functional MBL‐dependent LCP activation was associated with individual MBL2 haplotypes in individuals homozygous for null and replete haplotypes (p
ISSN:0892-6638
1530-6860
DOI:10.1096/fasebj.21.5.A181-d