Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses

Opitz, L.; Salaklang, Jatuporn; Büttner, Hermann; Reichl, Udo; Wolff, Michael W.

doi:10.1016/j.vaccine.2006.08.043

Cited by 67 publications

(34 citation statements)

References 24 publications

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“…The dsDNA measurements were done as described by Opitz et al (2007b) using the Quant-iT TM PicoGreen 1 dsDNA reagent from Molecular Probes Inc. (Cat #P7581, Eugene, OR). The assay was calibrated against lambda DNA (Cat.# D1501, Promega Corporation, Madison, WI) within the validated working range of 4-1,000 ng/mL (weighted regression; limit of detection: 0.66 ng/mL; limit of quantification: 2.36 ng/mL) using 100 mM citric acid buffer, pH 7.2, for dilutions.…”

Section: Dsdna-assaymentioning

confidence: 99%

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Wolff

Siewert

Lehmann

et al. 2009

Biotech & Bioengineering

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Smallpox is an acute, highly infectious viral disease unique to humans, and responsible for an estimated 300-500 million deaths in the 20th century. Following successful vaccination campaigns through the 19th and 20th centuries, smallpox was declared eradicated by the World Health Organization in 1980. However, the threat of using smallpox as a biological weapon prompted efforts of some governments to produce smallpox vaccines for emergency preparedness. An additional aspect for the interest in smallpox virus is its potential use as a platform technology for vector vaccines. In particular, the latter requires a high safety level for routine applications. IMVAMUNE, a third generation smallpox vaccine based on the attenuated Modified Vaccinia Ankara (MVA) virus, demonstrates superior safety compared to earlier generations and represents therefore an interesting choice as viral vector. Current downstream production processes of Vaccinia virus and MVA are mainly based on labor-intensive centrifugation and filtration methods, requiring expensive nuclease treatment in order to achieve sufficient low host-cell DNA levels for human vaccines. This study compares different ion exchange and pseudo-affinity membrane adsorbers (MA) to capture chicken embryo fibroblast cell-derived MVA-BN after cell homogenization and clarification. In parallel, the overall performance of classical bead-based resin chromatography (Cellufine sulfate and Toyopearl AF-Heparin) was investigated. The two tested pseudo-affinity MA (i.e., sulfated cellulose and heparin) were superior over the applied ion exchange MA in terms of virus yield and contaminant depletion. Furthermore, studies confirmed an expected increase in productivity resulting from the increased volume throughput of MA compared to classical bead-based column chromatography methods. Overall virus recovery was approximately 60% for both pseudo-affinity MA and the Cellufine sulfate resin. Depletion of total protein ranged between 86% and 102% for all tested matrices. Remaining dsDNA in the product fraction varied between 24% and 7% for the pseudo-affinity chromatography materials. Cellufine sulfate and the reinforced sulfated cellulose MA achieved the lowest dsDNA product contamination. Finally, by a combination of pseudo-affinity with anion exchange MA a further reduction of host-cell DNA was achieved.

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Section: Dsdna-assaymentioning

confidence: 99%

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Wolff

Siewert

Lehmann

et al. 2009

Biotech & Bioengineering

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“…Methods for virus purification include ultrafiltration [9,12] , size exclusion chromatography [7] , ion-exchange chromatography [11,22] and affinity chromatography [8,16,23] . In this study, a reliable and efficient method that combines tangential-flow ultrafiltration (TFF) and size-exclusion chromatography (SEC) is proposed for CSFV purification.…”

Section: Discussionmentioning

confidence: 99%

Efficient purification of cell culture-derived classical swine fever virus by ultrafiltration and size-exclusion chromatography

Wang¹,

Zhi²,

Guo³

et al. 2015

Front. Agr. Sci. Eng.

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Large-scale production of cell culture-based classical swine fever virus (CSFV) vaccine is hampered by the adverse reactions caused by contaminants from host cell and culture medium. Hence, we have developed an efficient method for purifying CSFV from cell-culture medium. Pure viral particles were obtained with two steps of tangential-flow filtration (TFF) and size-exclusion chromatography (SEC), and were compared with particles from ultracentrifugation by transmission electron microscopy (TEM), infectivity and recovery test, and real time fluorescent quantitative PCR (FQ-PCR). TFF concentrated the virus particles effectively with a retention rate of 98.5%, and 86.2% of viral particles were obtained from the ultrafiltration retentate through a Sepharose 4 F F column on a biological liquid chromatography system. CSFV purified by TFF-SEC or ultracentrifugation were both biologically active from 1.0Â10 -4.25 TCID 50 $mL -1 to 3.0Â10 -6.25 TCID 50 $mL -1 , but the combination of TFF and SEC produced more pure virus particles than by ultracentrifugation alone. In addition, pure CSFV particles with the expected diameter of 40-60 nm were roughly spherical without any visible contamination. Mice immunized with CSFV purified by TFF-SEC produced higher antibody levels compared with immunization with ultracentrifugation-purified CSFV (P < 0.05). The purification procedures in this study are reliable technically and feasible for purification of large volumes of viruses.

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“…Immobilized Euonymus europaeus lectin was an efficient affinity ligand used in the capture step for purification of human influenza A viruses derived from MDCK cells; the main targets were two viral glycoproteins (Opitz et al, 2007).…”

Section: Applications: Immobilized Lectins As Affinity Supports For Pmentioning

confidence: 99%

Protein Purification by Affinity Chromatography

Coelho¹,

Santos²,

Napoleão³

et al. 2012

Protein Purification

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Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses

Cited by 67 publications

References 24 publications

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Efficient purification of cell culture-derived classical swine fever virus by ultrafiltration and size-exclusion chromatography

Protein Purification by Affinity Chromatography

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