Residual HEK293 DNA is a critical impurity that must be minimized when generating recombinant AAV (rAAV) therapeutic products, as elevated levels can pose safety risks related to immunogenic and oncogenic effects. Residual host cell DNA quantification typically requires prior sample extraction to account for interference from complex biological matrices, including variations in buffer composition, residual proteins, and ionic strength. While extraction-based, off-line testing is adequate for final drug product testing, it limits real-time, in-process quality monitoring and optimization.

This application note delves into the development of the HostDetect™ HEK293 PCR DNA Quant Kit, showing how it has been designed to meet or exceed regulatory guidance by offering a PCR-based approach for quantifying residual host cell DNA. For HEK293, the high performing system is shown to deliver:

• Accelerated speed (< 2hrs)
• Greater sensitivity (detection as low as 0.03 pg/reaction)
• Higher precision (CV < 5% at 0.1 pg/reaction).

Guidelines are also provided for a streamlined, sample-extraction-free approach for residual DNA quantification using the HostDetect kit. A method that offers a timesaving, efficient, and practical solution to monitor residual HEK293 DNA during rAAV manufacturing, thereby facilitating GMP process optimization.

For research use only. Not for use in diagnostic procedures.