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A novel assay to quantitate MASP-2/ficolin-3 complexes in serum

Dorottya Csuka, Lea Munthe-Fog, Mikkel-Ole Skjoedt, Estrid Hein, Jakob T Bay, Lilian Varga, George Füst, Peter Garred

9 Citations (Scopus)

Abstract

Ficolin-1, -2 and -3 are recognition molecules in the lectin complement pathway and form complexes with serine proteases named MASP-1, -2 and -3 and two nonenzymatic proteins. MASP-2 is the main initiator of lectin pathway activation, while ficolin-3 is the most abundant ficolin molecule in the circulation. The significance of lectin pathway complexes in the circulation is unknown. Thus, we established an assay for the measurement of circulating MASP-2/ficolin-3 complexes. A quantitative sandwich ELISA was developed for the measurement of the MASP-2/ficolin-3 complexes in serum based on monoclonal antibodies against MASP-2 for coating and anti-ficolin-3 for detection. In addition, we assessed the serum concentrations of ficolin-3 and MASP-2 and the extent of ficolin-3 mediated C4 deposition on acetylated BSA in samples from 97 healthy donors. The median concentration of MASP-2/ficolin-3 complexes was found to be 119.7 AU/ml (range: 2.9-615.5 AU/ml). Significant correlations were found between the level of MASP-2/ficolin-3 complexes and the concentration of ficolin-3 (Spearman r=0.2532, p=0.0124), and MASP-2 (Spearman r=0.4505, p
Original languageEnglish
JournalJournal of Immunological Methods
Volume387
Issue number1-2
Pages (from-to)237-44
Number of pages8
ISSN0022-1759
DOIs
Publication statusPublished - 31 Jan 2013

Keywords

  • Antibodies, Monoclonal
  • Complement C4
  • Edetic Acid
  • Enzyme-Linked Immunosorbent Assay
  • Glycoproteins
  • Humans
  • Lectins
  • Mannose-Binding Protein-Associated Serine Proteases
  • Multiprotein Complexes
  • Polymorphism, Single Nucleotide
  • Reference Values
  • Serum

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