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European Biopharmaceutical Review

Mycoplasma Contamination - The Invisible Threat?

 

Cell cultures are increasingly being used for the manufacture of biopharmaceutical products. Contamination of these cell cultures by mycoplasma is a fundamental problem, as it potentially compromises the safety of the drugs being produced. Therefore, biopharmaceutical production must comply with different legal requirements and regulatory bodies all over the world. Recently, new techniques for rapid and comprehensive mycoplasma detection have been developed and validated in accordance with legal guidelines, replacing the time-consuming microbiology culture methods.

MYCOPLASMA CONTAMINATIONS IN BIOPHARMA

Contamination of cell cultures by mycoplasmas is a widespread and serious problem in biological research and biopharmaceutical production. Although testing cell cultures for mycoplasmas has increased in recent decades, it has been suggested that up to 30 per cent of cell cultures in current use are still infected, including those distributed by commercial culture collections (1).

Mycoplasmas (or Mollicutes) represent a distinct class of prokaryotes which are characterised by the lack of a substantial cell wall. With a size of only 0.2 to 2μm in diameter, they are considered to be the smallest bacteria known so far. Since the first report of the detection of mycoplasma in cell culture in the 1950s, more than 100 different mycoplasma species have been identified (2). The majority of the cell culture infections are caused by only a few species. Depending on the host cell line and the cell culture matrix used for cultivation, the species Mycoplasma arginini, M hyorhinis, M hominis, M orale, M fermentans and Acholeplasma laidlawii usually represent over 90 per cent of the identified contaminants. The frequency of these species may vary due to different applications and studies. If complex cell culture matrices are used for the production of biotechnological compounds, such as avian-, insect- or plant-based material, other mycoplasma species such as M gallisepticum, M synoviae or Spiroplasma citri may also be identified (3).


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Björn Breth, PhD, obtained his Masters degree in biotechnology working on proand eukaryotic fermentation processes and drug characterisation. During his PhD thesis, he focused on microarray-based analyses and characterisation of transcription factors. He continued his scientific career developing molecular test systems for high-throughput screening in cooperation with international biotechnology and pharmaceutical companies. At Greiner Bio-One GmbH, he is responsible for the development of bacterial, viral and human identification systems for the biopharmaceutical industry. Recently, he has become a member of the FDA mycoplasma working group.
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