Studies for the molecular basis of Mycoplasma infection and contamination, and the development of a novel rapid microbial method by metabolome analysis

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K16366
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47030:Pharmaceutical hygiene and biochemistry-related
• Research Institution: National Institute of Health Sciences
• Principal Investigator: Hayashi Katsuhiko 国立医薬品食品衛生研究所, 衛生微生物部, 研究員 (60804739)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: マイコプラズマ / メタボローム解析

Outline of Final Research Achievements

In this study, metabolome analysis was performed to detect mycoplasma contaminants in drug-producing cell culture using a mass spectrometer. Through this analysis, we tried to elucidate the mechanism of mycoplasma contamination.
When CHO-DG44 cells, which is used for antibody production, was infected with mycoplasma, mycoplasma proliferated without affecting the viability and proliferation of the cells. Even if we inoculated the amount of mycoplasma below the detection limit required for quality control of pharmaceuticals, the number of bacteria exceeded the number after cultivation. We analysed mycoplasma infected cell and culture supernatant by mass LC-MS/MS, resulting in the detection of lipid molecules specific for mycoplasma contamination.

Free Research Field

衛生微生物

Academic Significance and Societal Importance of the Research Achievements

医薬品の製造細胞が、マイコプラズマに汚染されると、医薬品の品質に問題が生じる可能性がある。マイコプラズマ汚染の検出には、多量の細胞が必要であり、細胞を使用しない検出法が求められている。
本研究の成果によって、わずかなマイコプラズマ汚染であっても、細胞培養後には影響のある菌量へと増殖すること、また、細胞培養の培地からマイコプラズマ汚染に特異的な脂質分子が検出されたことから、細胞を使用せずにマイコプラズマ汚染を検出する方法として利用できる可能性が示唆された。