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Application of Imaging Flow Cytometry for the Characterization of Intracellular Attributes in Chinese Hamster Ovary Cell Lines at the Single Cell Level

Pekle, Eva, Smith, Andrew, Rosignoli, Guglielmo, Sellick, Christopher, Smales, Christopher Mark, Pearce, Claire (2019) Application of Imaging Flow Cytometry for the Characterization of Intracellular Attributes in Chinese Hamster Ovary Cell Lines at the Single Cell Level. Biotechnology Journal, . Article Number 1800675. ISSN 1860-6768. (doi:10.1002/biot.201800675) (KAR id:73370)

Abstract

Biopharmaceutical manufacturing using Chinese hamster ovary (CHO) cells requires the

generation of high-producing clonal cell lines. During cell line development, cell cloning using

fluorescence activated cell sorting (FACS) has the potential to combine isolation of single cells

with sorting based on specific cellular attributes that correlate with productivity and/or growth,

identifying cell lines with desirable phenotypes for manufacturing. This study describes the

application of imaging flow cytometry (IFC) to characterize recombinant cell lines at the single

cell level to identify cell attributes predictive of productivity. IFC assays to quantify organelle

content, and recombinant heavy (HC) and light (LC) chain polypeptide and mRNA amounts in

single cells were developed. The assays were then validated against orthogonal standard flow

cytometry, western blot and qRT-PCR methods. We describe how these IFC assays may be

used in cell line development and show how cellular properties can be correlated with

productivity at the single cell level, allowing the isolation of such cells during the cloning

process. Our analysis found HC polypeptide and mRNA to be predictive of productivity early

in the culture, however specific organelle content did not show any correlation with

productivity.

Item Type: Article
DOI/Identification number: 10.1002/biot.201800675
Uncontrolled keywords: Chinese hamster ovary (CHO) cells, Single cell imaging, Imaging flow cytometry, organelles
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Mark Smales
Date Deposited: 04 Apr 2019 10:16 UTC
Last Modified: 28 Jul 2022 22:09 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/73370 (The current URI for this page, for reference purposes)

University of Kent Author Information

Smales, Christopher Mark.

Creator's ORCID: https://orcid.org/0000-0002-2762-4724
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