title: The isolation and identification of a secreted biomarker associated with cell stress in serum-free CHO cell culture.
abstract: One approach to improving mammalian culture productivity has been to reduce cell stress and cell death in the bioreactor, thus enhancing productivity through a longer phase of viability. Here we describe the isolation and identification of a biomarker for stress and viability loss in CHO culture. Using SELDI-TOF mass spectrometry to profile the protein component of supernatant culture media we have identified a peak at 7.7 kDa that was associated with loss of viability toward the end of the culture and simulated stress from both toxic metabolite accumulation and nutrient depletion. The relative intensity (signal/noise ratio) of the peak increased rapidly at the onset of dropping viability toward the end of the growth phase. Also, the peak height was seen to increase significantly when cells were grown under conditions emulating ammonia accumulation and glutamine deprivation. The species has been identified as a fragment of Galectin-1 (Gal-1) via MS/MS fingerprinting. We propose that this peak could be utilized as a marker for early onset of stress in cell culture. This work demonstrates the efficacy of SELDI technology to identify biomarkers in mammalian cell culture and highlights its value as a tool for the monitoring and improvement of culture processes.
authors: School of Chemical and Bioprocess Engineering, and Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors: John F
authors: JF
authors: Woolley
authors: Mohamed
authors: M
authors: Al-Rubeai
mesh_terms: D000818:Animals; D015415:Biomarkers; D016466:CHO Cells; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free / Q000737:chemistry*; D037483:Galectin 1 / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D008970:Molecular Weight; D013312:Stress, Physiological
chemical_list: D015415:Biomarkers; D016895:Culture Media, Serum-Free; D037483:Galectin 1