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Purification and Biochemical Characterization of Functional Complement Factor H From Human Plasma Fractions

Purification and Biochemical Characterization of Functional Complement Factor H From Human Plasma Fractions

H Brandstätter, P Schulz, I Polunic, C Kannicht, G Kohla, J Römisch

Vox Sang. 2012 Oct;103(3):201-12.

PMID: 22497541

Abstract:

Background and objectives:
Complement factor H (CFH) acts as major regulator of the alternative pathway of complement and its mutations and polymorphisms predispose to various human diseases. We aimed to develop a scalable purification process of CFH from human plasma fractions to supply a pathogen-safe and functional CFH concentrate.
Materials and methods:
Starting with intermediates of cold ethanol fractionation of plasma, CFH purification was performed with three chromatographic steps including solvent detergent treatment and nanofiltration. CFH functionality was tested by a haemolysis assay using sheep erythrocytes, by determining decay acceleration activity on C3 convertases and cofactor activity of C3b cleavage. CFH identity was confirmed by Western blot and mass spectrometry.
Results:
Three scalable chromatographic steps highly purified full-length and native CFH from human plasma fractions. The purification process enabled the removal of truncated and dysfunctional CFH species, yielding a native CFH concentrate as demonstrated in sensitive functional in vitro assays.
Conclusion:
This novel process provides a pathogen-safe and functional CFH concentrate that can be produced on an industrial scale and is suitable for pre-/clinical studies.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP80295654	Complement factor H from human plasma		80295-65-4	Price

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