Monoclonal antibody purification by ceramic hydroxyapatite chromatography

Area of Science:

• Biochemistry
• Chemical Engineering
• Biotechnology

Background:

• Monoclonal antibodies (Mab) are crucial therapeutics requiring high purity.
• Protein A chromatography is a common initial purification step for Mab.
• Further purification is often needed to remove residual impurities.

Purpose of the Study:

• To evaluate hydroxyapatite chromatography for Mab purification.
• To optimize lab-scale ceramic hydroxyapatite (CHT) methods for high Mab concentration.
• To achieve acceptable clearance of cell culture impurities using CHT.

Main Methods:

• Utilized ceramic hydroxyapatite (CHT) Type I, 40 μm chromatography.
• Employed two scouting methods for initial Mab purification.
• Optimized purification through linear phosphate screening, linear alkaline salt screening, and alkaline salt step gradients.

Main Results:

• Achieved high concentrations of purified Mab using optimized CHT methods.
• Demonstrated acceptable clearance of cell culture impurities.
• Milligram amounts of Mab were purified, typical for laboratory scale.

Conclusions:

• Hydroxyapatite chromatography is an excellent method for purifying Mab.
• Optimized CHT protocols enable high-concentration Mab purification at lab scale.
• CHT offers a viable downstream processing step for Mab therapeutics.