Poster Presentations
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Application of a HT magnetic bead based DNA extraction system to diverse mAb process intermediates
Ana Krstanovic-Anastassiades, Assistant Scientist, Merck Serono
The limit of 100 pg DNA per dose of a therapeutic protein set by regulatory authorities roughly equals the amount of DNA from less than 17 diploid Chinese hamster ovary (CHO) cells. To determine such trace amounts of DNA, the chosen analytical method must be extremely sensitive and robust.
Here, the evaluation and the qualification of a DNA extraction method on magnetic beads are described. The DNA extraction is aided by the use of an automated robotic platform, QIASymphony, prior to quantification by qPCR. This extraction method qualification constitutes an essence to support the process characterization and validation by QbD where the precision and the reproducibility of the analytical method will aid in shaping the production process design space which links CPPs to CQAs.
The study includes the tests designed to determine the total precision and accuracy of the extraction, the applicability to various matrices, and the repeatability of the extraction efficiency. Moreover, the range of linearity of the extraction procedure in regards to varying protein and DNA levels was also verified, to account for the range of concentrations of proteins and DNA which can be observed in process characterization studies.
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