This application note describes a method for detecting residual CHO host cell DNA using the Agilent Mx3005P QPCR System. The method uses sequence-specific primers and probe to detect CHO host cell DNA, producing more specific, sensitive, and faster results than conventional methods such as blot hybridization.
Chinese Hamster Ovary (CHO) cells are commonly used for producing human therapeutic proteins, necessitating sensitive detection of residual host cell DNA. This document presents an application note detailing the use of quantitative real-time polymerase chain reaction (qPCR) with the Agilent Mx3005P QPCR System to detect these contaminants.
The maximum acceptable amount of residual host DNA is 100 pg per therapeutic dose.
These assays produce results in less than three hours.
The conditions are 10 minutes at 95 °C followed by 40 cycles of 15 seconds at 95 °C and 1 minute at 60 °C.