A high-throughput approach to developing and optimizing mixed-mode membrane chromatography for protein purification.

Area of Science:

• Biotechnology
• Chemical Engineering
• Biopharmaceutical Manufacturing

Background:

• Membrane chromatography is an efficient alternative to packed-bed column chromatography for therapeutic protein purification.
• Key advantages of membrane chromatography include higher productivity and reduced buffer consumption.
• High-throughput screening (HTS) for membrane chromatography development has been challenging, hindering faster optimization.

Purpose of the Study:

• To develop a novel, high-throughput screening technique for optimizing membrane chromatography conditions.
• To enable rapid screening of multiple membrane chromatography conditions for monoclonal antibody (mAb) purification.
• To reduce development times and material requirements in biopharmaceutical purification processes.

Main Methods:

• A 96-well plate format was utilized for simultaneous screening of various membrane chromatography conditions.
• Four mixed-mode cation exchange membranes and one cation exchange membrane were evaluated.
• An incremental loading strategy (50 mg/ml increments up to 450 mg/ml) was employed, leveraging faster membrane mass transfer for reduced mixing times.

Main Results:

• The developed screening technique demonstrated significantly reduced mixing times (up to 60-fold) compared to resin-based methods.
• Screening results were comparable to larger-scale membrane purification runs.
• The technique allows for exploration of a wider operating space and requires substantially less feed material.

Conclusions:

• The novel 96-well plate screening method accelerates the optimization of membrane chromatography for monoclonal antibody purification.
• This HTS approach offers a significant advantage over traditional methods by reducing development timelines and resource needs.
• The technique is robust and provides reliable data for scaling up purification processes.