Using titer and titer normalized to confluence are complementary strategies for obtaining Chinese hamster ovary cell lines with high volumetric productivity of etanercept

Nuša Pristovšek, Henning Gram Hansen*, Daria Sergeeva, Nicole Borth, Gyun Min Lee, Mikael Rørdam Andersen, Helene Faustrup Kildegaard

*Corresponding author for this work

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Abstract

The selection of clonally-derived Chinese hamster ovary (CHO) cell lines with the highest production rate of recombinant glycoproteins remains a big challenge during early stages of cell line development. Different strategies using either product titer or product titer normalized to cell number are being used to assess suspension-adapted clones when grown statically in microtiter plates. However, no reported study so far has performed a direct head-to-head comparison of these two early reporters for predicting clone performance. Therefore, we developed a screening platform for high-throughput analysis of titer and confluence of etanercept-producing clones. We then performed an unbiased comparison of clone ranking based on either titer or titer normalized to confluence (TTC). Using two different suspension cultivation vessels, we demonstrate that titer- or TTC-based ranking gives rise to the selection of clones with similar volumetric productivity in batch cultures. Therefore, a combinatorial titer- and TTC-based ranking is proposed, allowing for selection of distinct clones with both, high integral viable cell density (IVCD) and high specific productivity features, respectively. This contributes to selection of a versatile panel of clones that can be further characterized and from which the final producer clone can be selected that best fits the production requirements.
Original languageEnglish
Article number1700216
JournalBiotechnology Journal
Volume13
Issue number3
Number of pages10
ISSN1860-6768
DOIs
Publication statusPublished - 2018

Keywords

  • CHO cells
  • Cell line development
  • High-throughput
  • Industrial Biotechnology
  • Recombinant proteins
  • Screening

Cite this

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title = "Using titer and titer normalized to confluence are complementary strategies for obtaining Chinese hamster ovary cell lines with high volumetric productivity of etanercept",
abstract = "The selection of clonally-derived Chinese hamster ovary (CHO) cell lines with the highest production rate of recombinant glycoproteins remains a big challenge during early stages of cell line development. Different strategies using either product titer or product titer normalized to cell number are being used to assess suspension-adapted clones when grown statically in microtiter plates. However, no reported study so far has performed a direct head-to-head comparison of these two early reporters for predicting clone performance. Therefore, we developed a screening platform for high-throughput analysis of titer and confluence of etanercept-producing clones. We then performed an unbiased comparison of clone ranking based on either titer or titer normalized to confluence (TTC). Using two different suspension cultivation vessels, we demonstrate that titer- or TTC-based ranking gives rise to the selection of clones with similar volumetric productivity in batch cultures. Therefore, a combinatorial titer- and TTC-based ranking is proposed, allowing for selection of distinct clones with both, high integral viable cell density (IVCD) and high specific productivity features, respectively. This contributes to selection of a versatile panel of clones that can be further characterized and from which the final producer clone can be selected that best fits the production requirements.",
keywords = "CHO cells, Cell line development, High-throughput, Industrial Biotechnology, Recombinant proteins, Screening",
author = "Nuša Pristovšek and Hansen, {Henning Gram} and Daria Sergeeva and Nicole Borth and {Min Lee}, Gyun and Andersen, {Mikael R{\o}rdam} and Kildegaard, {Helene Faustrup}",
year = "2018",
doi = "10.1002/biot.201700216",
language = "English",
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journal = "Biotechnology Journal",
issn = "1860-6768",
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TY - JOUR

T1 - Using titer and titer normalized to confluence are complementary strategies for obtaining Chinese hamster ovary cell lines with high volumetric productivity of etanercept

AU - Pristovšek, Nuša

AU - Hansen, Henning Gram

AU - Sergeeva, Daria

AU - Borth, Nicole

AU - Min Lee, Gyun

AU - Andersen, Mikael Rørdam

AU - Kildegaard, Helene Faustrup

PY - 2018

Y1 - 2018

N2 - The selection of clonally-derived Chinese hamster ovary (CHO) cell lines with the highest production rate of recombinant glycoproteins remains a big challenge during early stages of cell line development. Different strategies using either product titer or product titer normalized to cell number are being used to assess suspension-adapted clones when grown statically in microtiter plates. However, no reported study so far has performed a direct head-to-head comparison of these two early reporters for predicting clone performance. Therefore, we developed a screening platform for high-throughput analysis of titer and confluence of etanercept-producing clones. We then performed an unbiased comparison of clone ranking based on either titer or titer normalized to confluence (TTC). Using two different suspension cultivation vessels, we demonstrate that titer- or TTC-based ranking gives rise to the selection of clones with similar volumetric productivity in batch cultures. Therefore, a combinatorial titer- and TTC-based ranking is proposed, allowing for selection of distinct clones with both, high integral viable cell density (IVCD) and high specific productivity features, respectively. This contributes to selection of a versatile panel of clones that can be further characterized and from which the final producer clone can be selected that best fits the production requirements.

AB - The selection of clonally-derived Chinese hamster ovary (CHO) cell lines with the highest production rate of recombinant glycoproteins remains a big challenge during early stages of cell line development. Different strategies using either product titer or product titer normalized to cell number are being used to assess suspension-adapted clones when grown statically in microtiter plates. However, no reported study so far has performed a direct head-to-head comparison of these two early reporters for predicting clone performance. Therefore, we developed a screening platform for high-throughput analysis of titer and confluence of etanercept-producing clones. We then performed an unbiased comparison of clone ranking based on either titer or titer normalized to confluence (TTC). Using two different suspension cultivation vessels, we demonstrate that titer- or TTC-based ranking gives rise to the selection of clones with similar volumetric productivity in batch cultures. Therefore, a combinatorial titer- and TTC-based ranking is proposed, allowing for selection of distinct clones with both, high integral viable cell density (IVCD) and high specific productivity features, respectively. This contributes to selection of a versatile panel of clones that can be further characterized and from which the final producer clone can be selected that best fits the production requirements.

KW - CHO cells

KW - Cell line development

KW - High-throughput

KW - Industrial Biotechnology

KW - Recombinant proteins

KW - Screening

U2 - 10.1002/biot.201700216

DO - 10.1002/biot.201700216

M3 - Journal article

VL - 13

JO - Biotechnology Journal

JF - Biotechnology Journal

SN - 1860-6768

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ER -