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Purification and functional characterization of C4b-binding protein (C4BP)
1Department of Laboratory Medicine, Section of Medical Protein Chemistry, The Wallenberg Laboratory, Lund University, Skåne University Hospital, Malmö, Sweden.
Methods in Molecular Biology (Clifton, N.J.)
|November 13, 2013
Summary
We developed a purification method for C4b-binding protein (C4BP), a key complement system inhibitor. This method allows for assessing C4BP
Area of Science:
- Biochemistry
- Immunology
- Complement System
Background:
- C4b-binding protein (C4BP) is a large glycoprotein found in human plasma.
- C4BP is the primary regulator of the classical and lectin complement pathways.
- It controls complement reactions mediated by C4b.
Purpose of the Study:
- To describe a method for purifying C4BP from human plasma.
- To develop a functional assay for C4BP's cofactor activity.
Main Methods:
- Purification involved barium chloride precipitation, anion exchange chromatography, and gel filtration.
- Functional assessment utilized C4BP's cofactor role in Factor I-mediated C4b degradation.
Main Results:
- A reproducible method for C4BP purification from human plasma was established.
- A functional assay was developed to measure C4BP's cofactor activity.
Conclusions:
- The described purification method yields functional C4BP.
- The functional assay provides a means to assess C4BP's role in complement regulation.
