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With its leading CRO position in drug discovery, Creative Biolabs is exceptional in providing state-of-the-art drug discovery services to biotech and pharmaceutical customers worldwide to meet their preclinical, innovative drug discovery programs. Complement Factor H is an abundant plasma and serum protein that is an important regulator of the activity of the complement system. It is made up of a chain of twenty small homologous domains, called CCP (complement control protein) domains, each about 60 amino acids long, and contains two internal disulfide bridges. Here, we summarized the immobilized cardiolipin-based CFH purification protocol.
Immobilized Cardiolipin-Based CFH Purification Protocol
Cardiolipin is dissolved in ethanol and dried onto glass beads that are packed into a chromatography column. Plasma or serum is taken and diluted with water to reduce ionic strength, then poured through the column. Several proteins bind to the column (FH, variable amounts of IgG, IgM, IgA, beta2 glycoprotein 1, small quantity of C1q). These are eluted with high salt. IgG is removed by passage through a protein G column. FH is isolated to 95-99% purity by gel filtration on a Superose 6 column.
Fig.1 Flow chart of immobilized cardiolipin-based CFH purification protocol. (Creative Biolabs)
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Immobilized cardiolipin matrices leverage the high-affinity interaction between cardiolipin and CFH, enabling selective capture from complex biological samples. By covalently coupling cardiolipin to resin surfaces via amine-reactive chemistries, protocols achieve stable ligand attachment while preserving CFH-binding capacity. Buffer optimization—specifically, incorporating low-salt conditions with calcium chelators—minimizes nonspecific interactions, ensuring >90% purity post-elution. This approach effectively isolates functional CFH, as validated by SPR and cofactor activity assays. For large-scale applications, dynamic binding capacities exceeding 15 mg/mL resin support cost-efficient manufacturing.
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Creative Biolabs provides a range of complement testing services to meet the full range of early drug development needs of our customers. Our clients are in the pharmaceutical, biotechnology, and academic research fields. Our professional scientists and excellent technical support team provide you with full service. If you need more detailed information, please feel free to contact us.
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