In this study we have chosen to characterise the critical HCP components in terms of their persistence in a platform for MAb production. This is performed using 2D-polyacryamide gel electrophoresis and mass spectrometry analysis. Fed-batch cell culture and scale-down processing methodologies provide the basis to investigate the impact of both up and downstream process variables. This integrated approach is applied to null and producer cell lines. In one example we identify a number of intracellular proteins (e.g. protein disulphide isomerise; elongation factor 2; calreticulin) that exhibit a significant change in abundance with progression of culture relative to the general increase in HCP concentration.
Such characterisation is important for an understanding of process consistency and robustness, as the subsequent downstream process must be able to cope with such changes in relative abundance. Ultimately the information may enable a more directed approach to the development of purification strategies.
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