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Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells
Published on: January 29, 2014
Distribution and determinants of circulating complement factor H concentration determined by a high-throughput
Reecha Sofat1, P Patrizia Mangione, J Ruth Gallimore
1Centre for Clinical Pharmacology, Division of Medicine, University College London, Rayne Building, 5 University Street, London, WC1E 6JJ, UK.
Journal of Immunological Methods
|February 5, 2013
Summary
A new high throughput nephelometric assay for complement factor H (fH) is now available. This accurate and reliable assay reveals fH levels are linked to body fat and lipids in the general population.
Area of Science:
- Biochemistry
- Immunology
- Clinical Chemistry
Background:
- Research on complement factor H (fH) in human disease is limited by the absence of suitable assays for large-scale studies.
- A novel high throughput nephelometric assay for fH has been developed and validated.
Purpose of the Study:
- To develop and validate a high throughput nephelometric assay for human complement factor H (fH).
- To investigate the distribution and determinants of circulating fH concentration in a large human population.
Main Methods:
- Commercial radial immunodiffusion (RID) assay reagents were adapted for a Siemens BNII nephelometric instrument.
- Assay performance was evaluated against the commercial RID assay using healthy volunteer samples.
- Circulating fH concentration was analyzed in a large population sample to determine associations with lipids and body fat indices.
Main Results:
- The nephelometric assay demonstrated high recovery (~100%), reproducibility (intra-assay CV 11%, inter-assay CV 5-15%), and a wider operating range than the RID assay.
- Complement factor H was found to be a stable analyte, unaffected by freeze-thaw cycles or acute inflammatory stimuli.
- Population analysis revealed associations between plasma fH concentration, circulating lipids, and body fat indices.
Conclusions:
- The developed assay is the first high throughput method for measuring circulating fH, offering accuracy, reliability, and reproducibility.
- This assay provides crucial population-level data on fH distribution and its association with metabolic factors.
- The findings establish a valuable reference for future clinical and epidemiological research involving complement factor H.
