Cambio - Excellence in Molecular Biology

PCR

PCR : Rest DNA Testing

Detection of residual host cell DNA by dPCR with MiQuant® Residual DNA dPCR kits for E. coli, CHO or HEK-293:

  • Novel rapid, robust and sensitive method
  • Samples can be tested without performing DNA extraction
  • Meets EP, WHO and FDA criteria
  • Designed for use with the QIAGEN® QIAcuity® dPCR system
  • Aliquoted PCR mix ensures long-term stability
  • Lyophilized kit components simplify logistics and storage

 

Background & Description

Chinese hamster ovary cells (CHO), E. coli, and human embryonic kidney cells (HEK-293) have been widely used to manufacture recombinant proteins for human therapeutic use. After the synthesis of such immunotherapeutic or biosimilar, it is found that there could be some residual presence of DNA from the host cell. The World Health Organization (WHO), the Food and Drug Agency (FDA), and the European Pharmacopoeia (EP) recommend that the residual DNA in a manufactured final product needs to be less than 10 ng/dose. Even though downstream purification processes remove host cell residual DNA traces from products, testing for residual DNA is significant to ensure the customers’ safety.

The use of digital PCR (dPCR) is a very efficient way to determine the concentration of residual DNA in the final product. The MiQuant® Residual DNA dPCR kits for CHO, E. coli, and HEK-293 are the most accurate, efficient, and cost-effective way to analyze residual DNA in a sample without DNA extraction.

The kits are designed for QIAGEN®s QIAcuity® dPCR system as a novel rapid, robust, and sensitive method for absolute DNA quantification. This technology reduces the time needed for testing to less than three hours, including the dPCR sample preparation and thermocycling. Also, using these kits increases sensitivity since no DNA is lost during the extraction.

Short multicopy targets in the CHO (repeated element, ~ 1 million copies per genome), E. coli (7 genome copies), and HEK-293 (repeated element, ~ 1 million copies per genome) genomes are amplified. The length of the amplicons is < 200 bp and, therefore, short enough to allow the amplification of highly degraded DNA according to guidelines.

Moreover, each kit contains all the necessary PCR components, including Taq polymerase, primers, and dNTPs in a lyophilized mix, the Rehydration Buffer, dPCR Grade Water, and lyophilized Internal Control. The Internal Control is very reliable and will indicate any false negative result caused by PCR inhibition.

 

MiQuant® Residual DNA dPCR HEK-293

The MiQuant® Residual DNA dPCR HEK-293 is designed for the quantitative detection of residual DNA from HEK-293 cells in chemical and biological matrices. The MiQuant® Residual DNA dPCR HEK-293 is designed for use with the QIAGEN® QIAcuity® digital PCR (dPCR) system as a novel rapid, robust and sensitive method for the detection of residual host cell DNA.

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
58-0121MiQuant® Residual DNA dPCR HEK-29324 reactions £362.00 Quantity Add to Order
58-0122MiQuant® Residual DNA dPCR HEK-2934x 24 reactions POA Quantity Add to Order

Description

The MiQuant® Residual DNA dPCR HEK-293 is designed for the quantitative detection of residual DNA from HEK-293 cells in chemical and biological matrices.


The MiQuant® Residual DNA dPCR HEK-293 is designed for use with the QIAGEN® QIAcuity® digital PCR (dPCR) system as a novel rapid, robust and sensitive method for the detection of residual host cell DNA.

Type of PCR
Digital PCR (dPCR)

Recommended Use
To test product quality, the amount of residual DNA in a drug's final dosage form must follow guidelines established by the World Health Organization (WHO), the U.S. Food and Drug Administration (FDA), the European Pharmacopoeia, and other regulatory agencies.

 

 

Flyer


Kit Components
    dPCR HEK-293 Mix in aliquots of 24 reactions each
    Rehydration Buffer
    Positive Control DNA
    Internal Control DNA
    dPCR Grade Water

Package Size
Cat. No. 58-0121:
24 reactions

Cat. No. 58-0122:
4 x 24 reactions

Result Evaluation
Absolute Quantification

Required Consumables
    PCR reaction tubes
    PCR Clean™ (15-2025, 15-2001)
    PCR reaction tubes or plates
    MiQuant® dPCR Validation Standard HEK-293 (Cat. No. 58-0221)

Required Lab Device
    QIAcuity® digital PCR cycle
    QIAcuity® 24Well 26K Nanoplates
    Microcentrifuge for PCR reaction tubes
    Pipetting equipment

Shelf Life and Storage
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 store at ≤ -18°C for a maximum of one month.

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