Metagenomic characterization of parental and production CHO cell lines for detection of adventitious viruses

Viral contamination is a major concern for biological products. Therefore, virus testing of raw materials and cells is essential for the safety of the final product. We used high-throughput sequencing to detect viral-like sequences in selected CHO cell lines. Our aim was to test various approaches of sample preparation, to establish a pipeline for metagenomic analysis and to characterize standard viral metagenome of production and parental CHO cell lines. The comparison of the metagenomics composition of the differently prepared samples showed that among four tested approaches sequencing of ribosomal RNA depleted total RNA is the most promising approach. The metagenomics investigation of one production and three parental CHO cell lines of diverse origin did not indicate the presence of adventitious viral agents in the investigated samples. The study revealed an expected background of virus-like nucleic acids in the samples, which originate from remains of expression vectors, endogenized viral elements and residuals of bacteriophages.

Automated summary

This study utilized high-throughput sequencing to detect viral-like sequences in CHO cell lines, showing that sequencing of ribosomal RNA depleted total RNA is the most promising approach. No adventitious viral agents were found in the investigated cell lines, indicating a background of virus-like nucleic acids originating from expression vectors, endogenized viral elements, and residuals of bacteriophages.