Antibody purification from CHO cell supernatant using new multimodal membranes

Area of Science:

• Biotechnology
• Chemical Engineering
• Protein Purification

Background:

• Monoclonal antibodies (mAbs) are crucial therapeutics, requiring efficient purification from cell culture supernatants.
• Existing purification methods, like Protein A chromatography, have limitations for certain antibody types or process conditions.
• Chinese Hamster Ovary (CHO) cells are a common host for mAb production, necessitating effective downstream processing.

Purpose of the Study:

• To evaluate the efficacy of novel multimodal membranes (MMMs) for the capture step purification of monoclonal antibodies.
• To compare the performance of MMMs against commercial Protein A resin for human IgG1 (hIgG1) purification from CHO cell culture supernatant.
• To assess the impact of MMMs on product purity, host cell protein (HCP) levels, and elution conditions.

Main Methods:

• Development and application of multimodal membranes (MMMs) for antibody capture.
• Utilized a two-step chromatography process involving size-exclusion desalting followed by MMM chromatography.
• Analyzed product purity and host cell protein (HCP) levels using standard analytical techniques.
• Compared dynamic binding capacity and selectivity against a commercial Protein A resin.

Main Results:

• The MMM column exhibited higher dynamic binding capacity than Protein A resin at a 1-minute residence time.
• The two-step MMM process achieved high selectivity for hIgG1 capture from CHO supernatant.
• Product purity exceeded 98% with HCP levels below 20 ppm, comparable to Protein A purification.
• Elution at neutral pH was achieved, beneficial for preventing antibody aggregation, though it resulted in product dilution.

Conclusions:

• Multimodal membranes (MMMs) are highly effective for the capture step purification of proteins, including monoclonal antibodies.
• MMMs provide a viable alternative to Protein A chromatography, particularly for pH-sensitive mAbs or proteins lacking an Fc-binding domain.
• This purification strategy offers a valuable tool for downstream processing in biopharmaceutical manufacturing.