Applied Biosystems™ resDNASEQ™ Quantitative CHO DNA Kits

Description

• This quantitative PCR (qPCR)-based kit, part of the resDNASEQ™ Quantitative CHO DNA System, enables you to detect sub-picogram amounts of residual DNA from the Chinese hamster ovary (CHO) cell line in under 5 hours
• The system overcomes the limitations of traditional methods by combining high-recovery PrepSEQ™ sample preparation and TaqMan™-based quantitation of residual DNA
• Real-time quantitative PCR kit for highly sensitive residual CHO DNA quantitation • Easy to use, results in under 5 hours • Specific to CHO DNA, no cross-reactivity with unrelated DNA • Part of an integrated system that includes sample preparation, TaqMan™ assay and master mix, standard DNA, instrument, and software • Protocol available for automated sample preparation • Flexible sample throughput allows you to perform rigorous DNA clearance studies This kit is available with or without sample preparation reagents in the form of a PrepSEQ Residual DNA Sample Preparation Kit, which is optimized for highly efficient residual DNA recovery from complex mixtures of proteins, buffers, and salts
• Real-Time PCR for Highly Sensitive Quantitation The resDNASEQ™ Quantitative CHO DNA System provides highly sensitive detection of CHO DNA, allowing you to use small sample volumes to generate accurate results
• The broad linear range of TaqMan™ technology allows you to test samples with variable levels of CHO DNA in the same assay, such as in-process samples with higher amounts of DNA or bulk drug substance with very low amounts
• Figure 1 demonstrates the range and sensitivity of the assay
• Linearity is demonstrated by analysis of CHO cell standard DNA ranging from 0.3 ng to 3 fg
• Easy to Use, Results in Under 5 Hours First, you extract DNA from your in-process purification and drug substance samples
• Then you perform qPCR to compare DNA amounts in your test samples to a standard curve generated with known amounts of purified CHO cell standard DNA
• The easy workflow of the resDNASEQ™ Quantitative CHO DNA System consists of sample preparation, assay setup, and instrument run and data analysis—all of which you can perform in under 5 hours
• Specific to CHO DNA, No Cross-Reactivity With Unrelated DNA The target of the assay is highly specific, so that it only detects a hamster-specific region of a multicopy genetic element
• We selected this region using extensive bioinformatic analysis of multiple related and unrelated species
• Testing of assay performance confirmed that the assay is specific to hamster DNA and is unaffected by the presence of as much as 100 ng of unrelated DNA in a test sample
• Consistent Performance Even With Fragmented DNA For accurate quantitation of residual CHO DNA, assay results must be unaffected by the size of the DNA molecules present in the test sample
• To test the effect of DNA fragment size on assay performance, we fragmented high molecular weight CHO genomic DNA into low molecular weight DNA by sonication
• Figure 2 demonstrates that the threshold cycle (Ct) values for the reactions with the sonicated low molecular weight DNA were comparable to those of the undigested high molecular weight DNA
• These results demonstrate that consistent performance was obtained with the kit irrespective of DNA molecular weight.