Pyrogen detection methods: Comparison of bovine whole blood assay (bWBA) and monocyte activation test (MAT)

BMC Clinical Pharmacology

Manfred Kietzmann¹  Stephan Schumacher¹  Christian Wunderlich¹ 

[1] University of Veterinary Medicine Hannover, Foundation, Institute of Pharmacology, Toxicology and Pharmacy, Bünteweg 17, Hannover 30559, Germany

关键词: Lipoteichoic acid;    Lipopolysaccharide;    Monocyte activation test;    Pyrogen;    Prostaglandin E₂;    Bovine whole blood;    Endotoxin;   

Others  :  1084770
DOI  :  10.1186/2050-6511-15-50

 received in 2014-05-26, accepted in 2014-09-04,  发布年份 2014

【 摘 要 】

Background

Pyrogen detection is of utmost importance in pharmaceutical industry, laboratories and health care institutions. As an alternative to the animal-consuming rabbit pyrogen test or Limulus amoebocyte lysate test, the monocyte activation test was introduced as a gold standard method in the European Pharmacopoeia. However, the monocyte activation test has not gained wide acceptance in practice.

Methods

We stimulated bovine whole blood with different endotoxin preparations (lipopolysaccharide E.coli 0127:B8 and 0113:H10), as well as the non-endotoxin pyrogens peptidoglycan and lipoteichoic acid. Prostaglandin E₂ (PGE₂) served as read out.

Results

Employing PGE₂ as read out enabled detection limits of 0.04 EU/ml for lipopolysaccharide 0127:B8, 0.25 EU/ml for lipopolysaccharide 0113:H10 and 10 μg/ml of lipoteichoic acid as well as peptidoglycan. To evaluate the bWBA test system as a possible alternative to the MAT we performed a peer-to-peer comparison of the two methods and confirmed similar sensitivities.

Conclusions

In conclusion, the bovine whole blood assay (bWBA) reproducibly enabled sensitive detection of endotoxin and non-endotoxin pyrogens and may thus become a viable alternative for pyrogen testing.

【 授权许可】

   
2014 Wunderlich et al.; licensee BioMed Central Ltd.

【 图 表 】

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