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.