Multiple reaction monitoring targeted LC-MS analysis of potential cell death marker proteins for increased bioprocess control.
作者： Simone Albrecht , Christian Kaisermayer , David Reinhart , Monica Ambrose , Renate Kunert
DOI: 10.1007/S00216-018-1029-3
关键词:
摘要: The monitoring of protein biomarkers for the early prediction cell stress and death is a valuable tool process characterization efficient biomanufacturing control. A representative set six proteins, namely GPDH, PRDX1, LGALS1, CFL1, TAGLN2 MDH, which were identified in previous CHO-K1 model using discovery LC-MSE was translated into targeted liquid chromatography multiple reaction mass spectrometry (LC-MRM-MS) platform verified. universality markers confirmed growth three Chinese hamster ovary host lines (CHO-K1, CHO-S, CHO-DG44) grown batch culture two different types basal media. LC-MRM-MS also applied to spent media (n = 39) from four perfusion series. Stable isotope-labelled peptide analogues stable monoclonal antibody used improved quantitation simultaneous workflow reproducibility. Significant increases concentrations observed all viability marker proteins upon increased dead numbers allowed discrimination with densities below above 1 × 106 dead cells/mL highlights potential selected bioprocess
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