Genetic mutation analysis at early stages of cell line development using next generation sequencing
| Autor: | Mei Liu, Samantha Swahn, Helen McLaughlin, Chapman Wright, Scott Estes, Chongfeng Xu, Joost Groot, Chao Sun, Eric Zheng |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine media_common.quotation_subject CHO Cells Computational biology Biology medicine.disease_cause 01 natural sciences Mass Spectrometry DNA sequencing 03 medical and health sciences Cricetulus 010608 biotechnology medicine Animals Quality (business) Cells Cultured Selection (genetic algorithm) media_common Genetics Mutation business.industry Computational Biology High-Throughput Nucleotide Sequencing Sequence Analysis DNA 030104 developmental biology Biopharmaceutical Workflow Proof of concept Analytics business Biotechnology |
| Zdroj: | Biotechnology Progress. 32:813-817 |
| ISSN: | 8756-7938 |
| Popis: | A central goal for most biopharmaceutical companies is to reduce the development timeline to reach clinical proof of concept. This objective requires the development of tools that ensure the quality of biotherapeutic material destined for the clinic. Recent advances in high throughput protein analytics provide confidence in our ability to assess productivity and product quality attributes at early stages of cell line development. However, one quality attribute has, until recently, been absent from the standard battery of analytical tests facilitating informed choices early in cell line selection: genetic sequence confirmation. Techniques historically used for mutation analysis, such as detailed mass spectrometry, have limitations on the sample number and turnaround times making it less attractive at early stages. Thus, we explored the utility of Next-Generation Sequencing (NGS) as a solution to address these limitations. Amplicon sequencing is one such NGS technique that is robust, rapid, sensitive, and amenable to multiplexing, all of which are essential attributes for our purposes. Here we report a NGS method based upon amplicon sequencing that has been successfully incorporated into our cell line development workflow alongside other high-throughput protein analytical assays. The NGS method has demonstrated its value by identifying at least one Chinese hamster ovary (CHO) clone expressing a variant form of the biotherapeutic in each of the four clinical programs in which it has been utilized. We believe this sequence confirmation method is essential to safely accelerating the time to clinical proof of concept of biotherapeutics, and guard against delays related to sequence mutations. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:813-817, 2016. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text