Effect of cell retention techniques in Komagataella phaffii lab-scale continuous processes

Area of Science:

• Biotechnology
• Bioprocess Engineering
• Cell Culture Technology

Background:

• Perfusion technology is crucial for continuous biotherapeutic production, primarily in mammalian systems.
• Application of perfusion to alternative hosts like Komagataella phaffii is limited.
• Efficient cell retention is vital for perfusion bioreactor performance.

Purpose of the Study:

• To investigate and compare two lab-scale cell retention technologies for Komagataella phaffii continuous perfusion processes.
• To evaluate the performance of acoustic-based (AP) and membrane-based (MP) processes in terms of productivity, viability, and cell density.
• To assess the suitability of these technologies for yeast-based continuous biomanufacturing.

Main Methods:

• Development of an acoustic-based process using an acoustic cell separator (ACS).
• Development of a membrane-based process utilizing a vibrating membrane filtration (VMF) device.
• Continuous culture of Komagataella phaffii for scFv13R4 antibody fragment production over 8-9 days.

Main Results:

• Both AP and MP maintained high cell viability (>90%) and productivity over extended periods.
• AP achieved higher volumetric (50.63 mg L⁻¹ day⁻¹) and specific (1.09 mg g⁻¹ day⁻¹) productivities compared to MP (32.29 mg L⁻¹ day⁻¹ and 0.44 mg g⁻¹ day⁻¹).
• MP reached 100% separation efficiency with biomass up to 74.1 g L⁻¹ DCW, while AP achieved 98% efficiency with 55.1 g L⁻¹ DCW, showing selectivity for larger yeast cells.

Conclusions:

• Acoustic-based cell retention offers higher productivity in Komagataella phaffii perfusion processes.
• Membrane-based filtration achieves higher biomass concentration but with lower specific productivity.
• Both technologies are viable for K. phaffii continuous perfusion, with acoustic separation showing potential for enhanced yields.