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Use of novel cystine analogs to decrease oxidative stress and control product quality.

Valentine Chevallier1, Marvin Zoller2, Nadine Kochanowski2

  • 1UCB Nordic A/S, Upstream Process Sciences, Copenhagen, Denmark; Technical University of Denmark, Department of Biotechnology and Biomedicine, Kgs. Lyngby, Denmark.

Journal of Biotechnology
|December 29, 2020
PubMed
Summary
This summary is machine-generated.

Replacing l-cysteine with analogs like DACDM in cell culture media improves recombinant protein quality by reducing variants and coloration. This also boosts intracellular glutathione, aiding oxidative stress regulation.

Keywords:
CHO cellsCell culture mediaCharge variantsCysteineGlutathioneMonoclonal antibody

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Area of Science:

  • Biotechnology
  • Biopharmaceutical Manufacturing
  • Cell Culture Technology

Background:

  • Cell culture media require continuous optimization for high yield and product quality.
  • Instability of components like l-cysteine can negatively impact bioprocesses and lead to product microheterogeneity.
  • l-cysteine, while beneficial for recombinant protein production, can cause undesirable product variants.

Purpose of the Study:

  • To evaluate alternative cysteine and cystine analogs for cell culture media.
  • To identify replacements that reduce product variants and coloration without compromising productivity.
  • To investigate the impact of these analogs on cellular oxidative stress regulation.

Main Methods:

  • Assessment of N-acetyl-cysteine, s-sulfocysteine, N,N'-diacetyl-l-cystine, and N,N'-diacetyl-l-cystine dimethylester (DACDM).
  • Comparison of charge variants and recombinant protein coloration levels.
  • Measurement of intracellular glutathione pool to assess oxidative stress regulation.

Main Results:

  • Replacement of l-cysteine with analogs, particularly DACDM, significantly reduced charge variants.
  • DACDM demonstrated a positive impact on recombinant protein coloration levels.
  • DACDM increased the intracellular glutathione pool, indicating a role in oxidative stress management.

Conclusions:

  • Cystine analogs, especially DACDM, are effective replacements for l-cysteine in cell culture.
  • DACDM improves recombinant protein quality by minimizing microheterogeneity and coloration.
  • DACDM enhances cellular oxidative stress defense mechanisms through glutathione modulation.