Residual HEK293 DNA Detection qPCR Kit

The Residual DNA Detection qPCR Kit is designed for the sensitive detection of trace HEK293 DNA in a sample using quantitative PCR (polymerase chain reaction). By using a specific primer and probe set that targets the Alu elements in HEK293, this kit can achieve a detection limit as low as 1 fg/µl while minimizing background noise. Each kit provides enough 2x qPCR Master Mix (no dye) for reactions and includes a positive control, dilution buffer, and ROX reference dye for generating reliable standard curves. The manufacture of biological drugs, including vaccines, antibodies, and recombinant proteins, is regulated by agencies such as the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) to ensure that clinical drugs do not contain harmful impurities. Most biologicals are prepared using host cell systems such as human cells, insect cells, or bacterial cells, and carry-over of host cell DNA must be kept to a minimum as it could compromise patient safety. Therefore, manufacturers must ensure that the amount of residual host cell DNA is kept within safety levels defined by regulatory agencies. While specific requirements may vary depending on the host cell type and route of administration, these agencies generally recommend a maximum limit of 10 ng of host cell DNA per drug dose. To facilitate quality control, the Residual HEK293 DNA Detection qPCR Kit enables the detection of HEK293 DNA at concentrations as low as 1 fg/µl, with no cross-reactivity to rodent cell DNA. This specificity is achieved by targeting Alu elements. Alu elements are primate specific short-interspersed nuclear elements (SINEs), a class of retrotransposons derived from the 7SL RNA gene. Due to their successful propagation, the human genome now contains over one million copies of Alu elements. The primers and probe are designed against a consensus sequence from a human Alu subfamily, ensuring high sensitivity and specificity for human DNA. Figure 1. Principle of Probe-Based qPCR. Detection utilizes a 6-FAM-labeled probe paired with Black Hole Quencher-1 (BHQ-1). (A) In the intact probe, fluorescence is quenched via Förster resonance energy transfer (FRET). (B) During the extension phase, the 5'-3' nuclease activity of Taq polymerase cleaves the probe, separating the 6-FAM fluorophore from the quencher to generate a fluorescent signal. The absorbance and emission maxima of 6-FAM are 495 nm and 520 nm, respectively.