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Publication Detail
Analysis of fouling and breakthrough of process related impurities during depth filtration using confocal microscopy.
  • Publication Type:
    Journal article
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  • Authors:
    Parau M, Johnson TF, Pullen J, Bracewell DG
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  • Publication date:
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  • Journal:
    Biotechnology Progress
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  • Country:
    United States
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Titre improvement has driven process intensification in mAb manufacture. However, this has come with the drawback of high cell densities and associated process related impurities such as cell debris, host cell protein (HCP) and DNA. This affects the capacity of depth filters and can lead to carryover of impurities to protein A chromatography leading to early resin fouling. New depth filter materials provide the opportunity to remove more process related impurities at this early stage in the process. Hence there is a need to understand the mechanism of impurity removal within these filters. In this work, the secondary depth filter Millistak+ X0HC (cellulose and diatomaceous earth) is compared with the X0SP (synthetic), by examining the breakthrough of DNA and HCP. Additionally, a novel method was developed to image the location of key impurities within the depth filter structure under a confocal microscope. Flux, tested at 75, 100 and 250LMH was found to affect the maximal throughput based on the max pressure of 30psi, but no significant changes were seen in the HCP and DNA breakthrough. However, a drop in cell culture viability, from 87% to 37%, lead to the DNA breakthrough at 10% decreasing from 81 to 55L/m2 for X0HC and from 105 to 47L/m2 for X0SP. The HCP breakthrough was not affected by cell culture viability or filter type. The X0SP filter has a 30-50% higher max throughput depending on viability, which can be explained by the confocal imaging where the debris and DNA are distributed differently in the layers of the filter pods, with more of the second tighter layer being utilised in the X0SP. This article is protected by copyright. All rights reserved.
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