Cambio - Excellence in Molecular Biology

Pyrogen Testing

Pyrogen Testing: Pyrogen Testing

Pyrogen Testing

Complete Pyrogen testing by combining the power of digital PCR with Monocyte Activation Test (MAT)- Minerva Biolabs' NAT-MAT®.
  • Replacing the Rabbit Pyrogen Test
  • Ph. Eur. 2.6.30 compliant
  • Excellent reproducibility
  • Enhanced precision and Sensitivity
  • Highly automated workflow
  • Compatible with QIAcuity® dPCR platform
  • Time-to-Result: 1 day
  • Analysis of results with easy-to-use software

In the pharmaceutical industry, pyrogen detection is mandatory to avoid life-threatening fever reactions that can be induced by both microbial and non-microbial substances. Traditionally, this test relied on animal-based methods, like the rabbit pyrogen test (RPT). With increasing awareness of ethical concerns and growing demand for alternatives, the Monocyte Activation Test (MAT) was introduced into the European Pharmacopeia (EP) in 2010 – providing a human in vitro system to detect and quantify pyrogenic substances in pharmaceutical products. Since the RPT will be discontinued in 2026 in Europe, regulatory authorities currently encourage manufacturers to implement MAT in their QC system.

The MAT is based on the activation of monocytes by pyrogenic substances present in the sample. Upon exposure to potential pyrogens, monocytes undergo a signalling pathway resulting in secretion of pro-inflammatory cytokines, such as IL-1β, TNFα and IL-6.

Minerva Biolabs has developed a next generation MAT system that measures the gene expression of IL-1β and TNFα using digital PCR - the NAT-MAT®. The protocol is optimized for fast and reliable pyrogen detection and offers robust and highly sensitive results. The NAT-MAT® enables complete pyrogen testing and detects endotoxin as well as non-endotoxin pyrogens. The test can be conducted as in-process control and as final release testing of medicinal products according to Ph. Eur. 2.6.30. Analysis of the results can be done automatically by Minerva Biolabs´ easy-to-use analysis software.

 

 

Find more information on Minerva Biolabs' NAT-MAT® in our flyer.
Download Flyer

MiQuant® CAR-T Cell Lenti – dPCR

The MiQuant® CAR-T Cell Lenti – dPCR kit is intended for the determination of vector copy numbers (VCN) in transduced cells to determine the average copies of integrated lentiviral vectors per cell genome after ex vivo transduction via digital PCR (dPCR).

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
58-0701MiQuant® CAR-T Cell Lenti – dPCR35ul primer/probe mix £426.00 Quantity Add to Order
58-0702MiQuant® CAR-T Cell Lenti – dPCR140ul primer/probe mix £1,415.00 Quantity Add to Order

Description

This kit provides a specific primer/probe mix and offers an open-platform solution for use with various dPCR devices (QIAcuity®, ddPCR™, Quantstudio™ Absolute Q™). MiQuant® CAR-T Cell Lenti – dPCR works for all HIV-1 based lentivirus transduction systems.

Type of PCR

Digital PCR (dPCR)

Kit Components
  • MiQuant® Lenti Mix (lyophilized)
  • Positive Control DNA (lyophilized)
  • dPCR grade Water
Package Size

Cat. No. 58-0701: 35 µl primer/probe mix

Cat. No. 58-0702: 140 µl primer/probe mix

Since every dPCR device requires different PCR reaction volumes, the number of PCR reactions are shown in the following table.

Number of PCR reactions

Result Evaluation

Absolute Quantification

Required Lab Device and Consumables
  • Digital PCR device with corresponding consumables (plates or stripes)
    • QIAcuity®
    • Quantstudio™ Absolute Q™
    • QX600™ or QX200™ ddPCR™
  • Corresponding Mastermix (QIAcuity® Probe PCR Kit (4x), Absolute Q™ DNA Digital PCR Master Mix (5x), ddPCR™ Multiplex Supermix (4x) or ddPCR™ Supermix for Probes (2x))
  • MiQuant® SP Beads (sample preparation kit, Cat. No. 56-4010/-4100)
  • 1.5 ml reaction tubes (low-bind, DNA- and RNA-free)
  • Microcentrifuge for 1.5 ml PCR tubes
  • Vortex mixer
  • Pipettes with corresponding filter tips

Optional:

  • Restriction enzymes (EcoRI-HF and NruI-HF; 20 U/μl)
  • PCR Clean™ (Cat. No. 15-2025 and 15-2001)
Shelf Life and Storage

Shipment at ambient temperature. The expiry date of the unopened package is marked on the package label. 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.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

Recommended Use

 

MiQuant® CAR-T Cell Lenti – dPCR can be used for the characterization of cellular products and for in-process control as well as final QC release testing in cell therapy production processes. Besides these applications the MiQuant® CAR-T Cell Lenti – dPCR kit can also be used for determining VCN in cell culture samples in research and development and is applicable for human or other mammalian cells (such as mice and rats).

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200