Bioprocess: Robustness with Respect to Mycoplasma Species

Area of Science:

• Biopharmaceutical Manufacturing
• Microbial Contamination Control
• Process Engineering

Background:

• Mycoplasma contamination poses a significant risk to biopharmaceutical production.
• Previous studies demonstrated mycoplasma clearance by bioprocessing units using Acholeplasma laidlawii.

Purpose of the Study:

• To evaluate the efficacy of bioprocessing unit operations in clearing Mycoplasma orale and Mycoplasma arginini.
• To assess the robustness of these clearance mechanisms against different mycoplasma species.

Main Methods:

• Spike/removal studies using protein A chromatography, solvent/detergent treatment, low-pH hold, and UV-C irradiation.
• On-resin growth studies to monitor mycoplasma viability.
• Evaluation of virus-retentive filters for mycoplasma clearance.

Main Results:

• Protein A chromatography demonstrated significant clearance for M. orale and M. arginini, comparable to A. laidlawii, with some carryover.
• On-resin studies indicated mycoplasma die-off over time.
• Solvent/detergent treatment inactivated M. arginini but not fully M. orale.
• Low-pH hold and UV-C irradiation showed species-specific inactivation efficacy.
• Virus-retentive filters effectively cleared A. laidlawii.

Conclusions:

• Early bioprocessing steps and barrier technologies largely mitigate mycoplasma risk (M. orale, M. arginini).
• Certain unit operations may require enhanced conditions for hardier mycoplasma species.
• Established manufacturing practices ensure the safety of biotechnological products.