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Quantification of the pro-form of human complement component factor D (adipsin).

Maiken Lumby Henriksen1, Christian Nielsen2, Dennis Pedersen3

  • 1Dept. of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.

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|June 9, 2022
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Summary

A new ELISA assay quantifies pro-factor D, revealing 8-15% of this complement protein circulates in its precursor form. Blood collection methods impact measured levels.

Keywords:
AdipsinComplement systemELISAFactor DMASP-3Profactor D

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Area of Science:

  • Immunology
  • Biochemistry
  • Complement System Biology

Background:

  • Factor D (adipsin) is a serine protease crucial for complement system activation, particularly the alternative pathway.
  • It cleaves factor B, forming the C3 convertase C3bBb, central to complement amplification.
  • Adipocytes produce factor D as a pro-form, requiring cleavage by MASP-3 to become mature and active.

Purpose of the Study:

  • To develop and validate an ELISA for quantifying the pro-form of complement factor D.
  • To determine the circulating levels of pro-factor D in healthy individuals.
  • To investigate the influence of blood sampling procedures on factor D conversion.

Main Methods:

  • Development and validation of a specific ELISA for pro-factor D.
  • Assay validation included working range, intra/inter-assay coefficients of variation (CVs), and recovery rate.
  • Measurement of pro-factor D plasma concentrations in Danish blood donors.

Main Results:

  • The developed ELISA demonstrated a working range of 0.82-25 ng/ml with acceptable CVs and >90% recovery.
  • Median plasma concentration of pro-factor D in Danish blood donors was 134 ng/ml.
  • Pro-factor D constitutes 8-15% of total factor D in circulation, and blood sampling significantly affects measured levels.

Conclusions:

  • The study successfully established a validated ELISA for pro-factor D quantification.
  • A significant proportion of factor D circulates as a pro-form, with levels influenced by sample handling.
  • Further research is needed to understand the regulation and functional implications of pro-factor D and MASP-3 activation.