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Metagenomic characterization of parental and production CHO cell lines for detection of adventitious viruses
Biologicals ( IF 1.7 ) Pub Date : 2020-11-24 , DOI: 10.1016/j.biologicals.2020.11.001
Katarina Bačnik 1 , Denis Kutnjak 2 , Barbara Jerič Kokelj 2 , Nika Tuta 3 , Tan Lončar 3 , Matjaž Vogelsang 3 , Maja Ravnikar 4
Affiliation  

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.



中文翻译:

用于检测外来病毒的亲本和生产 CHO 细胞系的宏基因组表征

病毒污染是生物制品的主要问题。因此,原材料和细胞的病毒检测对于最终产品的安全至关重要。我们使用高通量测序来检测选定 CHO 细胞系中的病毒样序列。我们的目标是测试各种样品制备方法,建立宏基因组分析的管道,并表征生产和亲本 CHO 细胞系的标准病毒宏基因组。不同制备样品的宏基因组学组成的比较表明,在四种测试方法中,核糖体 RNA 耗尽的总 RNA 测序是最有前途的方法。对不同来源的一种生产和三种亲本 CHO 细胞系的宏基因组学研究并未表明所研究的样品中存在外来病毒因子。

更新日期:2020-11-24
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