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Detection of adventitious agents using next-generation sequencing.
Brenda Richards1, Sherry Cao2, Mark Plavsic3
1Functional Genomics & Science Computing; brenda.richards@genzyme.com.
PDA Journal of Pharmaceutical Science and Technology
|December 6, 2014
Summary
Next-generation sequencing effectively identifies bioreactor contaminants, including known and novel microbes. This sensitive method detects viruses and bacteria at very low levels, ensuring biopharmaceutical product safety.
Area of Science:
- Biotechnology
- Microbiology
- Genomics
Background:
- Biopharmaceutical manufacturing requires sterile conditions to prevent microbial contamination.
- Traditional methods for detecting contaminants are often limited to specific microorganisms.
- A sensitive and broad-range detection method is crucial for ensuring product safety.
Purpose of the Study:
- To evaluate next-generation sequencing (NGS) for identifying bioreactor contaminants.
- To establish the limits of detection for various microorganisms using NGS.
- To demonstrate the utility of NGS in detecting adventitious agents in biopharmaceutical production.
Main Methods:
- Utilized next-generation sequencing to analyze genomic data for microbial identification.
- Interrogated databases with sequencing data to identify potential adventitious agents.
- Performed spiking experiments with DNA viruses, RNA viruses, and bacteria in Chinese hamster ovary cells.
Main Results:
- Next-generation sequencing successfully identified the causative agent of a bioreactor contamination.
- All tested viruses were detected at less than 1 copy per cell.
- Bacteria were detected at a limit of 0.001 copy per cell.
Conclusions:
- Next-generation sequencing is a sensitive and valuable tool for detecting known and novel microbial contaminants in biopharmaceutical manufacturing.
- This approach addresses a critical need for broad-range microbial detection.
- NGS provides a complementary method to existing techniques for ensuring product quality and safety.
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