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DNA Removal for Bioprocessing Applications

e real-time PCR cycle thresholds (Ct) of residual lambda DNA detection using the conditions described in the text. Samples were amplified using an ABI 7900 real-time instrument. The Fold DNA Removal was calculated from the lambda DNA standard curve (slope=-3.3). Values are the average of replicate measurements. Duplicate real-time amplification curves for the samples presented in the table are also shown.


DNA Removal
Real-time PCR showed that treatment of live cells with 5 units of TURBO DNase reduced the amount of DNA contamination by 17.6 Ct, or more than 200,000 fold to 990 fg (Figure 1). Treatment with 5 units of wild type DNase I only reduced the DNA contamination by 11.5 Ct, or 2893-fold to 69 pg. Addition of 10-fold more of the wild type DNase enzyme reduced DNA content by 121,072-fold to 1,650 fgand was still not as effective as TURBO DNase used at 10 times lower unit concentration. TURBO DNase was able to reduce the copy number of DNA molecules from 1 billion to less than 5000 copies. In the best case, the competitors wild type DNase I, used at 10 times the unit concentration, reduced the DNA contamination to 8000 copies. This is an important finding, since clinical bioprocessing procedures often demand that the introduction of reagents, such as enzymes, into the process stream be restricted to the lowest effective level.

The superiority of TURBO DNase is striking when the comparison is based on the mass amount of enzyme added to the reacti
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