VMF of E. coli Cultures & Lysates

E. coli Harvest & Wash and Lysate Protein Clarification & Recovery using Vibro^® Membrane Filtration (VMF)

VMF performs better than conventional TFF for the processing of high‑density E. coli cultures and lysates. VMF demonstrated high protein recovery and solids handling at low TMP for three different expressed proteins, making it a robust, high-performance solution for demanding midstream applications. 

A “near-universal clarification platform”

To identify a “near-universal clarification platform” capable of handling diverse bioprocessing needs, this study compared the performance of Vibro^® Membrane Filtration (VMF), Hollow Fiber (HF) and Cassettes (CAS) in: 

• Cell harvest and wash of E. coli cells
• Lysate clarification and recovery of three different recombinant proteins

VMF outperformed the other modalities in both process steps, operating reliably at low TMP and achieving superior cell wash and target protein recovery compared to conventional TFF technologies like HF and CAS.

100% Cell Wash Efficiency with VMF

VMF was the most effective modality for cell wash, demonstrated by a measured 100% BSA clearance after three diafiltration volumes (DV). In contrast, HF only achieved ~90% clearance, while CAS was unable to complete three DV in the first place.

Superior Target Protein Recovery

Lysate protein recovery was tested across three different recombinant proteins of varying sizes. VMF achieved the best product recovery for all three proteins. For the largest protein (90-120 kDa) in particular, VMF more than doubled the product recovery (68%) compared to the next-best modality, HF (32%).

Robust operation at extremely low and uniform TMP

During both cell harvest and lysate clarification, VMF:

• Demonstrated solids handling capabilities on par with HF, whereas CAS loads consistently had to be reduced to allow operation
• Met the overall process targets
• Operated at a much lower and more uniform TMP compared to HF and CAS

Additionally, VMF’s unique process control allowed the flux to double by increasing the TMP during diafiltration, indicating a potential for further process optimization.