FUJIFILM Wako Pure Chemicals Launches New Pyrogen and Endotoxin Tests

OSAKA, Japan, June 27th, 2024 - FUJIFILM Wako Pure Chemicals Corporation announces the launch of two new tests: LumiMAT™ Pyrogen Detection Kit (LumiMAT), a next-generation monocyte activation test (MAT)*1 for in vitro*2 pyrogen testing, and PYROSTAR™ Neo+, a recombinant protein reagent for the detection of bacterial endotoxins. Alternatives to traditional pyrogen and endotoxin tests, the new LumiMAT and PYROSTAR™ Neo+ testing kits will be available worldwide in July 2024.

Pyrogens, including gram-negative bacterial endotoxins, are substances which produce fever, inflammatory reactions, and, in extreme cases, death,*³ when introduced into the bloodstream. Guidelines require that parenteral biological products, drugs, and medical devices that come in contact with the blood stream are tested for pyrogens.

Developed as an alternative to the rabbit pyrogen test, LumiMAT screens several cell lines*⁴ and is a rapid, robust, and highly sensitive assay for the detection of pyrogens, offering several advantages over traditional MAT assays currently available on the market. Most commercially available MAT assays utilize peripheral blood mononuclear cells (PBMCs)^*5 to measure cytokines released by monocytic cells upon exposure to pyrogens via ELISA,*⁶ and can result in lot-to-lot variation and long testing times (up to 1.5 days). In contrast, LumiMAT uses human NOMO-1 cells*⁷ selected in a screen based on their high sensitivity to lipopolysaccharide (LPS) and a wide range of non-endotoxin pyrogens, as well as their reproducibility and stability. In addition, LumiMAT utilizes a luciferase reporter assay*⁸ to detect activation of the transcription factor NF-κB,*⁹ producing results in approximately 5 hours. The European Pharmacopoeia recently announced plans to replace the rabbit pyrogen test by 2026, with a clear preference for the MAT.

PYROSTAR Neo+, a recombinant protein-reagent for the detection of gram-negative bacterial endotoxin utilizes the Kinetic Chromogenic Assay (KCA)*10 to measure endotoxin content. The reagent is composed of recombinant Factor C, Factor B, and a proclotting enzyme, allowing for endotoxin detection that offers the same sensitivity as the traditional LAL*¹¹ reagent. In addition, PYROSTAR Neo+ shows reduced interference with some drugs and improved reactivity to naturally occurring endotoxins, enabling use in a wider range of samples and applications, including product safety assessment and testing of pharmaceutical materials and water used for their manufacture.

said Koichi Yoshida, president of FUJIFILM Wako Pure Chemicals Corporation.

*1 A test to detect pyrogenic (fever-causing) substances based on the activation of human monocytes and the release of inflammatory cytokines in response to exogenous endotoxins and non-endotoxin pyrogens.
*2 A test performed on human or animal cells or tissues in a culture vessel or other medium.
*3 Solati S., Zhang T., Timman S. (2022). The monocyte activation test detects potentiated cytokine release resulting from the synergistic effect of endotoxin and non-endotoxin pyrogens. Innate Immun 28(3-4), pp. 130-137. PMID: 35491666.
*4 Cell lines are permanently established cells that can be stably grown and cultured for a long period of time.
*5 Mononuclear cells isolated from peripheral blood, including lymphocytes (T cells, B cells, NK cells) and monocytes. In humans, lymphocytes constitute the majority of the PBMC population.
*6 Enzyme-Linked Immuno-Sorbent Assay, an immunological assay commonly used to measure antibodies, antigens, proteins, and glycoproteins in biological samples using an enzymatic reaction.
*7 Human monocytic leukemia cell line.
*8 A test method used to determine if a substance can activate or repress the expression of a target gene. In the LumiMAT assay, pyrogenic substances are detected and quantified using reporter cells in which binding of pyrogenic agents to the Toll-like receptor (TLR) activates NF-κB and induces expression of luciferase protein.
*9 Nuclear Factor kappa B, a transcription factor that regulates the expression of genes involved in immune and inflammatory responses.
*10 A method for detecting endotoxin content, this assay measures color development that results from the breakdown of a substrate in the reagent caused by reaction with endotoxin.
*11 Limulus Amebocyte Lysate, a reagent for endotoxin assay.

LumiMAT™ uses NanoLuc® technology; NanoLuc® technology is licensed from Promega Corporation. Licensed Patents: U.S. Pat No. 8557970 and U.S. Pat. No. 8669103, and all patents and patents pending which claim priority to the same priority application(s) as U.S. Pat. No. 8557970 and U.S. Pat. No.8669103. NanoLuc® is a registered trademark of Promega Corporation.