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一种利用纳米流体学和光电定位技术的新型哺乳动物细胞系开发平台。

A novel mammalian cell line development platform utilizing nanofluidics and optoelectro positioning technology.

作者信息

Le Kim, Tan Christopher, Gupta Shivani, Guhan Trupti, Barkhordarian Hedieh, Lull Jonathan, Stevens Jennitte, Munro Trent

机构信息

Drug Substance Technologies, Process Development, Amgen Inc., Thousand Oaks, CA, 91320.

Attribute Sciences, Process Development, Amgen Inc., Thousand Oaks, CA, 91320.

出版信息

Biotechnol Prog. 2018 Nov;34(6):1438-1446. doi: 10.1002/btpr.2690. Epub 2018 Sep 19.

Abstract

Generating a highly productive cell line is resource intensive and typically involves long timelines because of the need to screen large numbers of candidates in protein production studies. This has led to miniaturization and automation strategies to allow for reductions in resources and higher throughput. Current approaches rely on the use of standard cell culture vessels and bulky liquid handling equipment. New nanofludic technologies offer novel solutions to surpass these limits, further miniaturizing cell culture volumes (10 times smaller) by growing cells on custom nanofluidic chips. Berkeley Lights' OptoElectro Positioning technology projects light patterns to activate photoconductors that gently repel cells to manipulate single cells on nanofluidic culturing chips. Using a fully integrated technology platform (Beacon), common cell culture tasks can be programmed through software, allowing maintenance and analysis of thousands of cell lines in parallel on a single chip. Here, we describe the ability to perform key cell line development work on the Beacon platform. We demonstrate that commercial production Chinese hamster ovary cell lines can be isolated, cultured, screened, and exported at high efficiency. We compare this process head to head with a FACS-enabled microtiter plate-based workflow and demonstrate generation of comparable clonal cell lines with reduced resources. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1438-1446, 2018.

摘要

生成一个高生产力的细胞系需要大量资源,并且由于在蛋白质生产研究中需要筛选大量候选细胞,通常耗时较长。这促使了小型化和自动化策略的出现,以减少资源消耗并提高通量。目前的方法依赖于使用标准细胞培养容器和笨重的液体处理设备。新的纳米流体技术提供了新颖的解决方案来突破这些限制,通过在定制的纳米流体芯片上培养细胞,进一步将细胞培养体积缩小(小10倍)。伯克利光公司的光电定位技术投射光图案来激活光电导体,从而轻柔地排斥细胞,以便在纳米流体培养芯片上操控单个细胞。使用一个完全集成的技术平台(Beacon),常见的细胞培养任务可以通过软件进行编程,从而能够在单个芯片上并行维护和分析数千个细胞系。在这里,我们描述了在Beacon平台上执行关键细胞系开发工作的能力。我们证明了可以高效地分离、培养、筛选和输出商业化生产的中国仓鼠卵巢细胞系。我们将这个过程与基于流式细胞仪的微孔板工作流程进行了直接比较,并证明了在减少资源的情况下能够产生可比的克隆细胞系。© 2018美国化学工程师学会生物技术进展,34:1438 - 1446,2018。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b93e/6585769/6e3700c4e3f6/BTPR-34-1438-g001.jpg

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