Murine leukemia virus clearance by flocculation and microfiltration

Akeprathumchai, Saengchai; Han, Binbing; Wickramasinghe, S. Ranil; Carlson, Jonathan O.; Czermak, Peter; Preibeta, Katrin

doi:10.1002/bit.20312

Cited by 14 publications

(7 citation statements)

References 30 publications

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“…Similar wall shear rates were used by Wickramasinghe et al (2004b) for tangential flow microfiltration of minute virus of mice. Higher wall shear rates have also been reported (Akeprathumchai et al, 2004). Since human influenza virus is known to be fragile and easily destroyed, the actual wall shear rate used in a manufacturing process should be sufficiently low to maximize isolation of intact virus particles and minimize damage to virus particles by shear forces during cross flow.…”

Section: Discussionmentioning

confidence: 97%

Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification

Wickramasinghe

Kalbfuss

Zimmermann

et al. 2005

Biotechnol. Bioeng.

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Large scale purification of viruses and viral vectors for gene therapy applications and viral vaccines is a major separation challenge. Here tangential flow microfiltration and ultrafiltration using flat sheet membranes has been investigated for concentration of human influenza A virus. Ultrafiltration membranes with molecular weight cutoffs of 100 and 300 kDa as well as 0.1, 0.2 and 0.45 microm microfiltration membranes have been tested. The results indicate that use of 300 kDa membranes not only concentrate the virus particles but also lead to a significant removal of host cell proteins and DNA in the permeate. Tangential flow filtration may be used to fractionate virus particles. Human influenza A virus particles are spherical with an average size of 100 nm. Use of a 0.1 microm membrane leads to passage of virus particles less than 100 nm into the permeate and an increase of larger particles in the retentate. These results suggest that control of the transmembrane pressure, membrane pore size and pore size distribution could enable isolation of intact virus particles from damaged virions. Isolation of the virus particles of interest from viral fragments and other particulate matter could result in simplification of subsequent purification steps. Larger pore size membranes such as 0.45 microm that allow the passage of all virus particles may be used to remove host cell fragments. In addition virus particles attached to these fragments will be removed. Careful selection of membrane morphology and operating conditions will be essential in order to maximize the benefit of tangential flow filtration steps in the purification of viral products from cell cultures.

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Section: Discussionmentioning

confidence: 97%

Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification

Wickramasinghe

Kalbfuss

Zimmermann

et al. 2005

Biotechnol. Bioeng.

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“…While results are cell culture and pH-dependent, reductions in DNA concentrations of 1-3 log 10 reduction value (LRV) and HCP levels of 0.2-0.8 LRV have been reported [16,20]. Viruses have also been shown to precipitate at acidic pH conditions, resulting in 3-4 LRV murine leukemia virus removal [22,23]. For pH-sensitive molecules, product recovery as low as 40% has been reported, though recovery of greater than 90% is typical [11,20].…”

Section: Key Termsmentioning

confidence: 96%

Industrial application of impurity flocculation to streamline antibody purification processes

Felo¹,

Kang

Hamzik³

et al. 2015

Pharmaceutical Bioprocessing

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Flocculation technologies offer significant benefits to industrial mammalian cell culture processes, including increased clarification efficiency, impurity removal and process simplification. In this paper, the application and performance of flocculation technologies employed in the harvest process of monoclonal antibody Chinese hamster ovary cell culture are reviewed. Attention has been placed on technologies enhancing the removal of cells, cellular debris and reduction in impurities while maintaining the antibody in the product stream. Many flocculants are systematically evaluated with respect to their mechanism of action, impact on downstream processing and product quality, and potential disadvantages. Practical considerations and future directions for application of flocculation in antibody manufacturing are discussed.

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“…Notably, positively charged flocculants, in particular polyamines (Peram et al, ), divalent cations (Romero et al, ; Shpritzer et al, ), chitosan (Riske et al, ), and polydiallyldimethylammonium chloride (PDADMAC) (McNerney et al, ; Zhao et al, ) have been shown to be successful in inducing flocculation as a result of interactions between the flocculant and the negatively charged surfaces of cells and cell debris. A key benefit of flocculation is a potential reduction in process related impurities such as HCP, host DNA, and viruses (Akeprathumchai et al, ). However, purification performance in this step is culture dependent (Roush and Lu, ) and it would be challenging to develop this step as an alternative to downstream polishing chromatography steps.…”

Section: Introductionmentioning

confidence: 99%

Development of a novel and efficient cell culture flocculation process using a stimulus responsive polymer to streamline antibody purification processes

Kang

Hamzik²,

Felo³

et al. 2013

Biotech & Bioengineering

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Recent advances in mammalian cell culture processes have significantly increased product titers, but have also resulted in substantial increases in cell density and cellular debris as well as process and product related impurities. As such, with improvements in titer, corresponding improvements in downstream processing are essential. In this study we have developed an alternative antibody harvest process that incorporates flocculation using a novel stimulus responsive polymer, benzylated poly(allylamine), followed by depth filtration. As tested on multiple antibodies, this process demonstrates high process yield, improved clearance of cells and cell debris, and efficient reduction of aggregates, host cell proteins (HCP) and DNA. A wide operating window was established for this novel flocculation process through design of experiments condition screening and optimization. Residual levels of impurities in the Protein A eluate were achieved that potentially meet requirements of drug substance and thus alleviate the burden for further impurities removal in subsequent chromatography steps. In addition, efficient clearance of residual polymer was demonstrated using a fluorescence tagged polymer in the presence of a stimulus reagent. The mechanism of HCP and aggregates removal during flocculation was also explored. This novel and efficient process can be easily integrated into current mAb purification platforms, and may overcome downstream processing challenges. Biotechnol. Bioeng. 2013;110: 2928–2937. © 2013 Wiley Periodicals, Inc.

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Murine leukemia virus clearance by flocculation and microfiltration

Cited by 14 publications

References 30 publications

Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification

Tangential flow microfiltration and ultrafiltration for human influenza A virus concentration and purification

Industrial application of impurity flocculation to streamline antibody purification processes

Development of a novel and efficient cell culture flocculation process using a stimulus responsive polymer to streamline antibody purification processes

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