The availability of rapid and accurate testing methods for contaminants and impurities that originate from biopharmaceutical manufacturing processes or from the host cell line is key to enabling in-process testing during manufacturing. The results from those tests can then be used as part of downstream process characterization to make fact-based, go/no-go forward processing decisions.
The AutoMate ExpressNucleic Acid Extraction System is an easy-to-use benchtop instrument that enables hands-free automation of the nucleic acid extraction process for contaminant and impurity testing. The instrument uses the established PrepSEQ nucleic acid extraction chemistry for quantitative recovery of DNA and RNA from diverse samples types, including cell culture, raw material (i.e., serum), and in-process downstream purification samples such as column purification or bulk drug substance (BDS) samples.
The AutoMate Express platform can be used to perform nucleic acid extraction of the common contaminants for mammalian cell culture manufacturing, i.e., mycoplasmas, MMV, and vesiviruses. Flexible protocols are available to isolate DNA and RNA from a single organism, or a 3-in-1 surveillance panel protocol can simultaneously extract mycoplasma, MMV, and vesivirus nucleic acid from a single sample. Figures 1 and 2 demonstrate recovery of DNA from spikes of DNA or of cells/viruses of mycoplasmas and MMV into typical cell culture matrices, extracted using the AutoMate Express system.
Figure 1. Detection of (A) mycoplasma DNA and (B) MMV DNA spiked into PBS (blue), fetal calf serum (black), CHO medium (gray), and conditioned CHO medium (brown). For each matrix, a recovery control (RC) was used to assess extraction efficiency.
Figure 2. Detection of DNA from (A) M. arginini and (B) MMV spiked into PBS (blue), fetal calf serum (black), CHO medium (gray), and CHO conditioned medium (brown). For each plate, control reactions were run with 500, 1,000, and 2,000 copies of the mycoplasma discriminatory positive control or MMV positive control in the real-time PCR assay.
Closed system with walk-away automation
The AutoMate Express platform automatically performs all steps for efficient nucleic acid extraction from contaminants and impurities in a closed system. The system uses PrepSEQ Express kits that contain sealed cartridges with extraction reagents, and plastics, including tips, and sample, lysis, and elution tubes. The workflow (Figure 3) has been designed for a completely hands-free operation enabling reproducible extraction efficiencies and reduced time-to-results. The small footprint of the instrument enables easy integration into an existing lab setup. Up to 13 samples can be processed in under an hour, and the eluates from the extraction can be used directly in real-time PCR-Seq analytics assays.
Figure 3. One-hour workflow for the AutoMate Express™ system.