Crozier, Jo-Ann Teresa Siobhan; (1993) Enhancement of product recovery in mammalian cell culture. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The objective of this doctoral study was the enhancement of the early stages of protein purification from mammalian cell culture, specifically the removal of cells, cell debris and other contaminants from large-scale cultures producing antibodies by the aggregative action of long-chain polyelectrolytes. To accomplish this, a detailed examination was made of the cell culture itself, and of the primary downstream processing stages. Assays were developed to monitor the levels of product and contaminants throughout the process, and a mass balance of the cell separation was produced. Having established current separation practice and efficiency, flocculants were screened on a small scale to assess their ability to aggregate mammalian cells and debris. Assays were used to establish the ability of the flocculants to remove contaminants such as nucleic acids and extraneous proteins while leaving antibody levels and activity unchanged. In order to assess the effect of adding flocculant to a large-scale cell culture, a scaled-down version of an industrial separation was performed. A theoretical efficiency index was used to compare two centrifuges, and assays were performed to evaluate the effectiveness of the flocculant when exposed to conditions similar to those found on an industrial scale. Conclusions were drawn on the desirability of using such chemicals to assist in the early purification of large-scale cultures of mammalian cells.

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