Mycoplasma testing
Cell cultures used in the manufacture of biopharmaceuticals must be tested for Mycoplasma contamination. Mycoplasma contamination does not produce turbidity, but can have adverse effects on cell line characteristics/growth rates and virus production. Certain Mycoplasma species are also classed as human pathogens therefore their presence in a manufacturing process is of serious concern.
International regulatory guidelines vary slightly in their recommended detection methods.Variations of the following assays are used, all with multiple positive controls.
Mammalian cell culture |
The sample is incubated with monkey kidney cells and is then fixed, stained with a DNA-binding fluorochrome (Hoechst Stain), and evaluated microscopically by epifluorescence for the presence of Mycoplasma. |
Broth and agar |
Broth flasks are inoculated with the test article and are incubated aerobically. Samples from the broth flasks are sub-cultured on appropriate days onto agar plates. Plates are incubated under appropriate microaerophilic, anaerobic and aerobic conditions for a period of 7-14 days depending on the regulatory guidance being followed. |
Plate agar assays |
The sample is inoculated directly on agar plates and incubated under appropriate microaerophilic, anaerobic and aerobic conditions depending on the regulatory guidance being followed. Plates are examined for Mycoplasma no sooner than 14 days post-inoculation. |
Mycoplasmastasis. Mycoplasmastasis is described in the EP guidance, and represents a qualification test to examine the test sample for product-specific inhibitors that can interfere with the growth of Mycoplasma. Normally this test would be required only once for any given product provided there is no change in the manufacturing process.
Regulatory Compliance
All our Mycoplasma assays at a minimum meet the required regulatory standards.
- European Pharmacopoeia (EP 2.6.7 Mycoplasmas)
- US Code of Federal Regulations (21 CFR 610.30)
- “Characterization and Qualification of Cell Substrates and Other Biological Starting Materials Used in the Production of Viral Vaccines for the Prevention and Treatment of Infectious Diseases”
US Food and Drug Administration and the Center for Biologics Evaluation and Research (FDA/CBER) 2006 - Draft
- ‘Points to Consider in the characterization of cell lines used to produce biologicals.’
US Food and Drug Administration and the Center for Biologics Evaluation and Research (FDA/CBER) 1993
We also offer Mycoplasma testing in accordance with Good Laboratory Practice (GLP).