Mycoplasma Clearance and Risk Analysis in a Model Bioprocess

Area of Science:

• Biotechnology
• Microbiology
• Pharmaceutical Manufacturing

Background:

• Mycoplasmas are cell wall-deficient bacteria that can contaminate cell cultures.
• Their small size allows passage through 0.2 μm filters, posing a patient safety risk in biotechnology manufacturing.
• Undetected mycoplasma contamination can compromise the safety and efficacy of biopharmaceuticals.

Purpose of the Study:

• To evaluate the efficacy of standard monoclonal antibody (mAb) purification processes in clearing and inactivating mycoplasmas.
• To assess the removal and killing capabilities of protein A chromatography, low-pH holds, and solvent/detergent treatments.
• To ensure the safety of biotechnology-derived medicines by addressing potential mycoplasma contamination.

Main Methods:

• Spike/removal studies using *Acholeplasma laidlawii* as a model mycoplasma.
• Evaluation of protein A column chromatography for mycoplasma clearance.
• Assessment of low-pH hold steps (glycine and acetate buffers) for mycoplasma inactivation.
• Analysis of solvent/detergent treatment for mycoplasma killing efficacy.

Main Results:

• Protein A chromatography achieved significant mycoplasma clearance (4-5 log10 reduction).
• Column regeneration effectively prevented mycoplasma carryover between runs.
• Low-pH hold steps inactivated *A. laidlawii* (LRV ≥4.57), and solvent/detergent treatment achieved complete inactivation (LRV ≥5.95).

Conclusions:

• Typical monoclonal antibody purification operations effectively clear and inactivate mycoplasmas.
• These manufacturing steps provide critical control points for ensuring the microbiological safety of biopharmaceuticals.
• The findings enhance confidence in the safety assurance of biotechnology medicines for patients.