Highly Sensitive and Robust Automated DNA Extraction from Pharma Manufacturing In-Process Quality Control (QC) Samples using the MagNA Pure LC 2.0 System
ثبت نشده
چکیده
To ensure final product safety, clearance of host cell DNA from the drug substance is essential in the manufacturing process for therapeutic proteins and monoclonal antibody (MAB) drugs. We have developed and validated an automated process for monitoring clearance of Chinese Hamster Ovary (CHO) DNA impurities during the capture and polishing steps of downstream processing. This method, which eliminates manual steps in DNA extraction and subsequent qPCR analyses, can help overcome the analytics bottleneck during process development and routine testing. DNA extraction with Roche’s MagNA Pure LC System offers excellent DNA recovery rates and robustness towards matrix effects for subsequent highly sensitive quantitative PCR. The MagNA Pure LC System eliminates manual dilution of high protein or DNA loads, manual neutralization of samples from acidic protein purification, and manual addition of carrier RNA. Here, we show validation data from in-process control QC samples collected during the pharmaceutical manufacturing process that were spiked with defi ned amounts of CHO DNA, and from in-process controls, using Roche’s MagNA Pure LC System and Qiagen’s QIAcube Instrument. Highly Sensitive and Robust Automated DNA Extraction from Pharma Manufacturing In-Process Quality Control (QC) Samples using the MagNA Pure LC 2.0 System September 2011
منابع مشابه
Use of the MagNA pure LC automated nucleic acid extraction system followed by real-time reverse transcription-PCR for ultrasensitive quantitation of hepatitis C virus RNA.
Hepatitis C virus (HCV) infection is an increasing health problem worldwide. Quantitative assays for HCV viral load are valuable in predicting response to therapy and for following treatment efficacy. Unfortunately, most quantitative tests for HCV RNA are limited by poor sensitivity. We have developed a convenient, highly sensitive real-time reverse transcription-PCR assay for HCV RNA. The assa...
متن کاملHigh-throughput detection of bacterial, fungal and viral nucleic acids in routine microbiological sample types using one generic Pathogen Universal Protocol on the MagNA Pure 96 System
The objective of this study was to compare the MagNA Pure 96 Pathogen Universal protocol to the MagNA Pure LC DNA Isolation Kit III that is well established in our microbiological laboratory. For this we isolated nucleic acids from ten different sample materials: body fl uids (CSF, urine, sputum), feces, swabs, as well as whole blood, EDTA-, Citrate-plasma and serum. This spectrum is typical fo...
متن کاملComparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory
PURPOSE The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use. MATERIALS AND METHODS Venous blood samples from 22 healthy volunteers were analyzed using QIAamp Blood Mini Kit (Qiagen), MagNA Pure LC Nucleic Acid Isolation Ki...
متن کاملUse of an Automated Method Improves the Yield and Quality of Cell- Free Fetal DNA Extracted from Maternal Plasma To the Editor: Cell-free fetal DNA in the maternal circulation holds great potential for noninvasive prenatal diagnosis
To the Editor: Cell-free fetal DNA in the maternal circulation holds great potential for noninvasive prenatal diagnosis (1, 2). However, the manual isolation of cell-free DNA from plasma can be a time-consuming process, and current techniques involving spin columns usually require multiple reloadings of the columns with sample material, increasing the risk of contamination (3–5). Use of an auto...
متن کاملFully automated, internally controlled quantification of hepatitis B Virus DNA by real-time PCR by use of the MagNA Pure LC and LightCycler instruments.
We report on the development of a fully automated real-time PCR assay for the quantitative detection of hepatitis B virus (HBV) DNA in plasma with EDTA (EDTA plasma). The MagNA Pure LC instrument was used for automated DNA purification and automated preparation of PCR mixtures. Real-time PCR was performed on the LightCycler instrument. An internal amplification control was devised as a PCR comp...
متن کامل