Residual Host Cell 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
• Fast workflow as no sample preparation is required prior to PCR analysis
• Meets EP, WHO and FDA criteria
• Compatible with QIAcuity® dPCR platform
• Lyophilized kit components simplify logistics and storage

Residual host cell DNA testing is a critical aspect of quality control in the production of biopharmaceuticals, including recombinant proteins, monoclonal antibodies, and cell-based therapies. Host cell DNA refers to the genetic material originating from the cell line used to produce the biopharmaceutical product. For this purpose, CHO, HEK-293 and E. coli cells are frequently used. Even after extensive downstream purification processes, trace amounts of residual host cell DNA may remain in the final product, posing potential safety and regulatory concerns.

The WHO, FDA, and the European Pharmacopoeia recommend that the residual DNA concentration is less than 10 ng/dose in the final product.

Minerva Biolabs has developed a digital PCR assay for the detection of residual host cell DNA for E.coli, CHO or HEK-293 - MiQuant® Residual DNA dPCR. The use of dPCR reduces time-to-result since no sample preparation is required prior to dPCR analysis and enables absolute quantification without a standard curve. With the help of the dPCR Validation Standards it is possible to determine a matrix-specific conversion factor that converts [copies/µl] into [pg/µl] to provide a quantification of the residual DNA in conformity to regulatory guidelines.

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.

The kits are designed for the QIAcuity® dPCR system and contain all necessary components, including Taq polymerase, primers, probes and dNTPs in a lyophilized mix, the rehydration buffer, dPCR grade water, and a lyophilized Internal Control that will indicate false negative results caused by PCR inhibition.