Adsorption of plasmid DNA on ceramic hydroxyapatite chromatographic materials.

The adsorption of plasmid DNA onto two different types of ceramic hydroxyapatite beads with a particle diameter of 20  μm, namely Ceramic Hydroxyapatite Type II and the Type III, which is not commercially available, were investigated. Type II and the Type III have a pore diameter of 80 and 240  nm, respectively. Equilibrium and dynamic binding capacity for a 4.

Matrix-assisted refolding of autoprotease fusion proteins on an ion exchange column: a kinetic investigation.

Matrix-assisted refolding is an excellent technique for performing refolding of recombinant proteins at high concentration because aggregation during refolding is partially suppressed. The autoprotease N(pro) and its engineered mutant EDDIE can be efficiently refolded on cation-exchangers. In the current work, denatured fusion proteins were loaded at different column saturations (5 and 50 mg mL(-1) gel), and refolding and self-cleavage were initiated during elution.

Matrix-assisted refolding of autoprotease fusion proteins on an ion exchange column.

Refolding of proteins must be performed under very dilute conditions to overcome the competing aggregation reaction, which has a high reaction order. Refolding on a chromatography column partially prevents formation of the intermediate form prone to aggregation. A chromatographic refolding procedure was developed using an autoprotease fusion protein with the mutant EDDIE from the N(pro) autoprotease of pestivirus.