Identification and characterization of Chinese hamster ovary (CHO) host cell protein (HCP) impurities by proteomic techniques can aid bioprocess design, leading to more efficient development and improved biopharmaceutical manufacturing operations.  Extracellular CHO HCP recovery for proteomic analysis is particularly challenging due to the relatively low protein concentration and complex media composition.  We have developed optimized protocols that improve proteome capture as a route to increasing the utility of proteomic methods, and this presentation discusses conditions that maximize extracellular CHO HCP detection for gel-based and shotgun workflows.  Twelve precipitation protocols were screened for protein recovery and optimized for a subset of the precipitants by design of experiment approaches.  Precipitation conditions were compared by two-dimensional electrophoresis and liquid chromatography coupled with mass spectrometry, with optimized recovery shown to differ between the two proteomic methods.  The optimized precipitation conditions were also applied to HCP characterization in common bioprocess streams to demonstrate their utility.

Optimizing Chinese hamster ovary host cell protein recovery for proteomic analysis

245th National Meeting (2013)

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