Large-scale methacrylate monolithic columns: design and properties

Podgornik, Aleš; Jančar, Janez; Merhar, M.; Kozamernik, S.; Glover, D.; Čuček, Karmen; Barut, Miloš; Štrancar, Aleš

doi:10.1016/j.jbbm.2004.01.002

Cited by 69 publications

(36 citation statements)

References 16 publications

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“…For the preparation of large volume tubular devices, a few annular shaped monoliths with different and well controlled inner and outer diameters are prepared and fitted one into another. These tubular methacrylate based columns with a volume of up to 8 L operating in radial direction at a maximal flow-rate of 10 L/min are produced by BIA Separations under the trade name CIMm Tube Monolithic Columns [91].…”

Section: Different Formats Of Monolithsmentioning

confidence: 99%

Preparation of methacrylate monoliths

Влах

Tennikova²

2007

J of Separation Science

150

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Rigid macroporous polymers developed in the early 1990s are widely used as efficient stationary phases for all types of chromatographic separations. The main advantages of so-called monolithic supports are their high hydraulic permeability and the dominance of the convection over the diffusion mechanism of mass-exchange under dynamic conditions that allow the separation to be carried out at extremely high flow rates and, consequently, during very short operation times. Among other types of macroporous polymers, the methacrylate-based monolithic materials represent the most popular and successfully explored class of sorbents. This review is an attempt to collect together the contributions of different groups working in the area of monolith preparation. Examples of different methcrylate monomers and crosslinkers, as well as porogenic solvents, including polymer ones, used in monolith preparation are discussed.

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Section: Different Formats Of Monolithsmentioning

confidence: 99%

Preparation of methacrylate monoliths

Влах

Tennikova²

2007

J of Separation Science

150

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“…During prolonged usage, preparative methacrylate monolithic columns have to be regenerated, commonly by using 1 M NaOH [8]. This treatment can be performed without any influence on the column, however, it is always questionable whether all the compounds attached to the matrix have been removed.…”

Section: Resultsmentioning

confidence: 99%

“…To follow the idea of short monolithic layers [4,5] they were designed in the format of tubes and operate in radial chromatographic mode [3, 6±8]. Despite this mode of operation, the complete usage of the monolith bed is achieved through extremely fast mass exchange [9] and chromatographic properties were shown to be flow-unaffected [3,8]. These performances were utilized for fast separation and purification of lignin peroxidases [10] and plasma proteins [6,11,12], for which easy scale-up of the chromatographic method was demonstrated [12].…”

Section: Introductionmentioning

confidence: 99%

“…The superior properties of monolithic columns for pDNA purification lead to their implementation in the industrial purification process of plasmid DNA intended for human gene therapy [14]. This process requires that the columns should not only exhibit outstanding properties in terms of efficiency and productivity [13] combined with high mechanical and chemical stability as well as regeneration capability with 1 M NaOH [8], but also be extensively character-ized before as well as throughout the usage to ensure a constant quality of the purified product.…”

Section: Introductionmentioning

confidence: 99%

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Noninvasive Methods for Characterization of Large‐Volume Monolithic Chromatographic Columns

et al. 2005

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Chromatographic monoliths exhibit several properties like flow-unaffected resolution and dynamic binding capacity, low pressure drop and high capacity for extremely large molecules, which are advantageous to the purification of large biomolecules such as proteins, DNA or even viruses. Recently, large-volume monolithic columns were introduced enabling the monoliths to be incorporated into industrial downstream processes. Due to the monolithic structure, however, conventional methods for resin characterization cannot be applied to monoliths, which limit their usage under cGMP conditions. In this article, a methodology of how to overcome this problem and to determine the most important parameters is presented. Several fast and noninvasive methods for monolith characterization are introduced. Their usage enables accurate determination of monolith pore size distribution and specific surface area, uniformity of the monolith as well as type and density of the active groups before or during monolith usage.

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“…This affects plasmid retention, binding capacity, purification yield or productivity and general economics of the purification process. 7,8 This study explores a cost-effective non-toxic scalable technique for rapid pDNA production employing a methacrylate monolithic adsorbent. The synthesis and characterization of the polymeric resin with a pore diameter distribution and structure tailored for pDNA binding and retention are presented.…”

Section: Introductionmentioning

confidence: 99%

Towards the design of a scalable and commercially viable technique for plasmid purification using a methacrylate monolithic stationary phase

Danquah

Forde

2007

J of Chemical Tech & Biotech

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A monolithic stationary phase was prepared via free radical co-polymerization of ethylene glycol dimethacrylate (EDMA) and glycidyl methacrylate (GMA) with pore diameter tailored specifically for plasmid binding, retention and elution. The polymer was functionalized with 2-chloro-N,N-diethylethylamine hydrochloride (DEAE-Cl) for anion-exchange purification of plasmid DNA (pDNA) from clarified lysate obtained from E. coli DH5α-pUC19 culture in a ribonuclease/protease-free environment. Characterization of the monolithic resin showed a porous material, with 68% of the pores existing in the matrix having diameters above 300 nm. The final product isolated from a single-stage 5 min anion-exchange purification was a pure and homogeneous supercoiled (SC) pDNA with no gDNA, RNA and protein contamination as confirmed by ethidium bromide agarose gel electrophoresis (EtBr-AGE), enzyme restriction analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This non-toxic technique is cGMP compatible and highly scalable for production of pDNA on a commercial level.

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Large-scale methacrylate monolithic columns: design and properties

Cited by 69 publications

References 16 publications

Preparation of methacrylate monoliths

Preparation of methacrylate monoliths

Noninvasive Methods for Characterization of Large‐Volume Monolithic Chromatographic Columns

Towards the design of a scalable and commercially viable technique for plasmid purification using a methacrylate monolithic stationary phase

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