Cell harvesting by cross-flow microfiltration using a shear-enhanced module

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Separation Science

Background:

• Recombinant protein production in bacteria like Vibrio cholerae is crucial for various applications.
• Efficient separation of target proteins from fermentation broth is a key challenge in bioprocessing.
• Conventional microfiltration methods can face limitations in protein recovery and flux.

Purpose of the Study:

• To evaluate a new mechanically agitated (rotational) shear filter for separating recombinant Vibrio cholerae protein.
• To assess the filter's performance in terms of protein recovery, permeate flux, and transmembrane pressure.
• To determine the filter's efficiency during concentration and diafiltration steps.

Main Methods:

• Utilized a DMF(TM) rotational shear filter for cross-flow microfiltration of bacterial fermentation broth.
• Employed a process of sixfold concentration followed by twofold diafiltration.
• Monitored transmembrane pressure, permeate flux, and protein transmission throughout the separation process.
• Evaluated filter cleaning efficacy using an enzymatic cleaning agent.

Main Results:

• Achieved higher protein recovery (97%) and permeate flux compared to common cell harvesting methods.
• Maintained a constant flux of 150 L/m(2) h with only a 10 kPa transmembrane pressure increase.
• Demonstrated high protein transmission: ~100% initially, >90% post-concentration, and >80% during diafiltration.
• Observed no significant decrease in pure water flux after enzymatic cleaning, indicating good filter cleanability.

Conclusions:

• The DMF(TM) rotational shear filter offers superior performance for Vibrio cholerae protein separation.
• The method enables efficient concentration and diafiltration with high protein recovery and stable operation.
• This technology presents a promising advancement for bioseparation processes in the pharmaceutical and biotechnology industries.