NAT-MAT® All-in-One System

NAT-MAT® All-in-One System provides all necessary components, including cells, extraction system, and dPCR assay to perform monocyte activation test (MAT) in order to detect pyrogen contaminations in pharmaceuticals.

The NAT-MAT® system relies on the use of NAT-MAT® Cells which are stimulated with a pyrogen standard and the sample to be tested followed by nucleic acid extraction (NAT-MAT® Extraction) and subsequent dPCR analysis of the gene expression of IL-1β and TNF-α (NAT-MAT® dPCR). By simultaneously measuring the gene expression of a housekeeping gene, the pyrogenic load can be normalized to the cell number, which allows highly accurate results.

Components

• NAT-MAT® Cells (Cat. No. 58-0501)
• NAT-MAT® Extraction (Cat. No. 58-0502)
• NAT-MAT® dPCR (Cat. No. 58-0503)

Package Size

Cat. No. 58-0511: 96 reactions

Result Evaluation

Absolute Quantification

Required Lab Device and Consumables

NAT-MAT® Cells:

• Liquid nitrogen or ≤-150 °C ultra-freezer
• CO² Incubator
• Laminar airflow cabinet
• Water bath
• Centrifuge (for 15 ml falcons)
• 15 ml falcon tubes
• Fetal bovine serum
• Inverted microscope (optional)
• Reference standard endotoxin (RSE), e.g. preparations from USP or EP (BRP)
• Sterile, non-pyrogenic water (LAL water)
• 2 relevant non-endotoxin pyrogens (optional)
• Pyrogen-free glass tubes
• Microcentrifuge
• Vortex mixer
• Pipettes with corresponding filter tips (10 μl, 100 μl, and 1000 μl)
• Multi-channel pipette (optional)

NAT-MAT® Extraction:

• 1-propanol, ≥99,5 %, p.a.
• Automated extraction device (e.g. KingFisher™ Flex)
• 96-well plates for automated extraction device
• Important: For manual extraction (not recommended): thermal shaker and magnetic rack

NAT-MAT® dPCR:

• QIAcuity® digital PCR system
• QIAcuity® Nanoplate 8.5k 96-well
• QIAcuity® OneStep Advanced Probe Kit
• Nanoplate tray
• Nanoplate seal
• 1.5 ml reaction tubes
• Microcentrifuge for 1.5 ml PCR tubes
• Vortex mixer
• Pipettes with corresponding filter tips
• Pipetting robot (optional, e.g. QIAgility® with nanoplate adapter)

Shipment, Shelf Life, and Storage

The expiry date of the unopened package is marked on the package label.

NAT-MAT® Cells: Shipment with dry ice (cells) and ambient temperature (cell culture media and plate) in 2 separate packages. The cells must be stored at ≤-150 °C or liquid nitrogen until use. The cell culture media must be stored at +2 to +8°C until use.

NAT-MAT® Extraction: Shipment and storage at ambient temperature.

NAT-MAT® dPCR: Shipment at ambient temperature. The kit components must be stored at +2 to +8°C until use. The rehydrated components must be stored at ≤ -18°C for a maximum of one month.