Mycoplasma: Characteristics and Implications in Biotechnology

What are Mycoplasma and why are they problematic in cell culture?

Mycoplasma are a genus of bacteria belonging to the class Mollicutes, distinguished by their lack of a cell wall, making them inherently resistant to beta-lactam antibiotics such as penicillin. Their minimal genome and small size (0.2–0.3 µm) allow them to pass through standard filters used in sterilisation. In cell culture, Mycoplasma contamination can alter cell metabolism, growth rates, and gene expression without obvious morphological changes, leading to unreliable experimental data and compromised biopharmaceutical production. Contamination is often undetected for prolonged periods, making routine screening essential.

How are Mycoplasma detected and prevented in biotech processes?

Detection methods include PCR-based assays, DNA staining (e.g., DAPI), enzyme-linked immunosorbent assays (ELISA), and culture techniques. PCR is widely preferred due to its sensitivity and speed. Prevention involves strict aseptic technique, regular testing of cell lines and reagents, and the use of Mycoplasma-free media and supplements. Regulatory guidelines from organisations such as the FDA, EMA, and ISO 11135 require routine Mycoplasma testing for cell-based therapies and biologics. In compliance with GMP and ICH Q5A, manufacturers must validate their processes to ensure Mycoplasma absence.

Related concepts

Mycoplasma contamination poses a critical risk in biopharmaceutical development, necessitating robust quality control. Regular testing using validated methods such as PCR or ELISA, combined with adherence to standards like ISO 11135 and USP <1111>, ensures product safety and regulatory compliance.