Inhibitory effects of persistent apoptotic cells on monoclonal antibody production in vitro: simple removal of non-viable cells improves antibody productivity by hybridoma cells in culture

Christopher D. Gregory, John D. Pound, Andrew Devitt, Megan Wilson-Jones, Parthasarathi Ray, Ruth J. Murray

Research output: Contribution to journalArticle

Abstract

Cells undergoing apoptosis in vivo are rapidly detected and cleared by phagocytes. Swift recognition and removal of apoptotic cells is important for normal tissue homeostasis and failure in the underlying clearance mechanisms has pathological consequences associated with inflammatory and auto-immune diseases. Cell cultures in vitro usually lack the capacity for removal of non-viable cells because of the absence of phagocytes and, as such, fail to emulate the healthy in vivo micro-environment from which dead cells are absent. While a key objective in cell culture is to maintain viability at maximal levels, cell death is unavoidable and non-viable cells frequently contaminate cultures in significant numbers. Here we show that the presence of apoptotic cells in monoclonal antibody-producing hybridoma cultures has markedly detrimental effects on antibody productivity. Removal of apoptotic hybridoma cells by macrophages at the time of seeding resulted in 100% improved antibody productivity that was, surprisingly to us, most pronounced late on in the cultures. Furthermore, we were able to recapitulate this effect using novel super-paramagnetic Dead-Cert Nanoparticles to remove non-viable cells simply and effectively at culture seeding. These results (1) provide direct evidence that apoptotic cells have a profound influence on their non-phagocytic neighbors in culture and (2) demonstrate the effectiveness of a simple dead-cell removal strategy for improving antibody manufacture in vitro.
LanguageEnglish
Pages370-376
Number of pages7
JournalMAbs
Volume1
Issue number4
DOIs
Publication statusPublished - Jul 2009

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Hybridomas
Antibody Formation
Cell Culture Techniques
Monoclonal Antibodies
Antibodies
Phagocytes
In Vitro Techniques
Immune System Diseases
Nanoparticles
Homeostasis
Cell Death
Macrophages
Apoptosis

Keywords

  • Antibodies, Monoclonal
  • Apoptosis
  • Cell Culture Techniques
  • Cell Survival
  • Humans
  • Hybridomas
  • Protein Engineering

Cite this

Gregory, Christopher D. ; Pound, John D. ; Devitt, Andrew ; Wilson-Jones, Megan ; Ray, Parthasarathi ; Murray, Ruth J. / Inhibitory effects of persistent apoptotic cells on monoclonal antibody production in vitro : simple removal of non-viable cells improves antibody productivity by hybridoma cells in culture. In: MAbs. 2009 ; Vol. 1, No. 4. pp. 370-376.
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Inhibitory effects of persistent apoptotic cells on monoclonal antibody production in vitro : simple removal of non-viable cells improves antibody productivity by hybridoma cells in culture. / Gregory, Christopher D.; Pound, John D.; Devitt, Andrew; Wilson-Jones, Megan; Ray, Parthasarathi; Murray, Ruth J.

In: MAbs, Vol. 1, No. 4, 07.2009, p. 370-376.

Research output: Contribution to journalArticle

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