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Accelerated clone selection for recombinant CHO CELLS using a FACS-based high-throughput screen.

Christine T DeMaria1, Victor Cairns, Cordula Schwarz

  • 1Therapeutic Protein Expression Group, Department of Cell & Protein Therapeutics, Genzyme Corporation, Framingham, Massachusetts 01701, USA. christine.demaria@genzyme.com

Biotechnology Progress
|January 31, 2007
PubMed
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This study introduces a novel flow cytometry method using a nonfluorescent reporter protein for rapid screening of high-producing cell clones. This technique efficiently identifies stable cell lines for therapeutic protein production.

Area of Science:

  • Biotechnology
  • Cell Biology
  • Protein Expression

Background:

  • Developing stable recombinant cell lines is crucial for therapeutic protein production.
  • Current screening methods can be time-consuming and may not efficiently identify high-producing clones.
  • A need exists for rapid and accurate methods to select cell lines with high therapeutic protein expression levels.

Purpose of the Study:

  • To develop a rapid and accurate method for identifying high-producing recombinant cell lines using flow cytometry.
  • To establish a screening process that allows for early-stage elimination of unstable clones.
  • To enable efficient selection of cell lines for therapeutic protein manufacturing.

Main Methods:

  • Coexpression of a therapeutic protein with a nonfluorescent reporter protein in CHO cells.

Related Experiment Videos

  • Linking therapeutic and reporter protein genes via an Internal Ribosome Entry Site (IRES) for co-transcription.
  • Utilizing flow cytometry with fluorescently labeled antibodies for reporter protein detection and clone screening.

    • Main Results:

    • The expression level of the reporter protein accurately correlated with the therapeutic protein expression level due to common mRNA.
    • Rapid and accurate screening of clones in 96-well plates was achieved.
    • Early-stage elimination of unstable clones was facilitated, improving cell line development efficiency.

    Conclusions:

    • This flow cytometry-based reporter system provides an effective process for generating recombinant cell lines with high therapeutic protein yields.
    • The method offers rapid clone screening and early elimination of unstable clones, streamlining cell line development.
    • The system's independence from therapeutic protein-specific antibodies allows for broad applicability across various cell line development processes.