SMB Operation for Three-Fraction Separations: Purification of Plasmid DNA

Paredes, Galatea; Mazzotti, Marco; Stadler, Joachim; Makart, Stefan; Morbidelli, Massimo

doi:10.1007/s10450-005-6033-1

Cited by 17 publications

(9 citation statements)

References 11 publications

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“…Most of them relate to separation problems that are pseudo-ternary separations, that is, where at least one of the compounds is baselineseparated from the others (Gottschlich and Kasche, 1997;Paredes et al, 2005). The advantages of SMB over batch chromatography are represented by a better productivity and stationary phase utilization.…”

Section: Introduction Mcsgp For Biomolecule Separationmentioning

confidence: 99%

Chromatographic separation of three monoclonal antibody variants using multicolumn countercurrent solvent gradient purification (MCSGP)

Müller‐Späth

Aumann

Melter

et al. 2008

Biotech & Bioengineering

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116

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Multicolumn countercurrent solvent gradient purification (MCSGP) is a continuous chromatographic process developed in recent years (Aumann and Morbidelli, 2007a; Aumann et al., 2007) that is particularly suited for applications in the field of bioseparations. Like batch chromatography, MCSGP is suitable for three-fraction chromatographic separations and able to perform solvent gradients but it is superior in terms of solvent consumption, yield, purity, and productivity due to the countercurrent movement of the liquid and the solid phases. In this work, the MCSGP process is applied to the separation of three monoclonal antibody variants on a conventional preparative cation exchange resin. The experimental process performance was compared to simulations based on a lumped kinetic model. Yield and purity values of the target variant of 93%, respectively were obtained experimentally. The batch reference process was clearly outperformed by the MCSGP process.

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Section: Introduction Mcsgp For Biomolecule Separationmentioning

confidence: 99%

Chromatographic separation of three monoclonal antibody variants using multicolumn countercurrent solvent gradient purification (MCSGP)

Müller‐Späth

Aumann

Melter

et al. 2008

Biotech & Bioengineering

Self Cite

116

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“…For SMB, CIP was proven to work for the separation of nucleosides and plasmid DNA (16,17). However, designed for two-fraction separations and a single-step gradient, SMB is only of limited use for the purification of biomolecules from a mixture of impurities with similar adsorptive properties.…”

Section: Mcsgp For Biomolecule Separationmentioning

confidence: 99%

Role of Cleaning-in-Place in the Purification of mAb Supernatants Using Continuous Cation Exchange Chromatography

Müller‐Späth

Aumann

Morbidelli

2009

Separation Science and Technology

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Monoclonal antibodies (mAbs) are among the most important therapeutic proteins for the treatment of cancer. They also find wide application in the field of diagnostics. The market for mAbs was US$ 5.4 billion in 2002 and is expected to triple by 2010 (1). MAbs are mainly produced in cell culture and purified by chromatography. Apart from the mAb, the downstream processing is confronted with a multitude of impurities from the upstream process. Some of these impurities bind strongly to the chromatographic stationary phase and can only be removed by separate cleaning solutions. Depending on the type of media used, e.g. affinity or ion exchange, differing cleaning regimes may have to be applied. In addition, sanitization is required to prevent microbial contamination. Both steps are closely tied to the mAb production step and need to be repeated in regular intervals.In this work, the impact of irreversible impurity adsorption on the separation performance is investigated experimentally; first for a batch column and then for a continuous chromatographic process. At first, the retention time of a model substance is established as a suitable measure to describe the degree of irreversible adsorption. The impact of the presence and the absence of cleaning-in-place (CIP) on the retention time of the model substance is demonstrated.Since the issue of column cleaning is particularly important if processes are operated in continuous mode, in the second part of this work the introduction of a CIP step in the continuous multicolumn countercurrent solvent gradient purification (MCSGP) process is investigated. The process purifies a mAb from a cell culture supernatant that also contains irreversibly adsorbing impuritiesor impurities that desorb in a later cycle of the process as a contamination. Following the same approach as for the batch experiments, the MCSGP process is operated in the presence and the absence of CIP in order to track prospective changes of the process performance caused by changes in the product retention time. Instead of the model substance, a cell culture supernatant, providing a real case impurity profile in an industrial application is used for the process. Although an impact of the product retention time change on the process yield is not observed it is shown that CIP is required for a stable long-term operation in terms of purity and system pressure drop. With the introduction of a CIP step the process was successfully operated for 9000 min (6 days) without interruption.

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“…The new challenge for the SMB technology is its application to the separation and purification of biomolecules [202]. Examples of products that are considered for SMBC separation and purification are proteins [203,204], amino acids [205], antibodies [206], nucleosides [207], and plasmid DNA [208].…”

Section: Thin-layer Chromatography (Tlc)mentioning

confidence: 99%

Separation Strategies for Processing of Dilute Liquid Streams

Mandal

Kulkarni

2011

International Journal of Chemical Engineering

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Processing of dilute liquid streams in the industries like food, agro-, biotechnology, pharmaceuticals, environment, and so forth needs special strategy for the separation and purification of the desired product and for environment friendly disposal of the waste stream. The separation strategy adopted to achieve the goal is extremely important from economic as well as from environmental point of view. In the present paper we have reviewed the various aspects of some selected universal separation strategies such as adsorption, membrane separation, electrophoresis, chromatographic separation, and electroosmosis that are exercised for processing of dilute liquid streams.

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SMB Operation for Three-Fraction Separations: Purification of Plasmid DNA

Abstract: The Simulated Moving Bed technology is extended to incorporate a cleaning in place step, and it is then applied by exploiting size exclusion chromatography to purify plasmid DNA. Experimental performances are discussed in the light of our theoretical understanding of the SMB behavior.

Cited by 17 publications

References 11 publications

Chromatographic separation of three monoclonal antibody variants using multicolumn countercurrent solvent gradient purification (MCSGP)

Chromatographic separation of three monoclonal antibody variants using multicolumn countercurrent solvent gradient purification (MCSGP)

Role of Cleaning-in-Place in the Purification of mAb Supernatants Using Continuous Cation Exchange Chromatography

Separation Strategies for Processing of Dilute Liquid Streams

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