Noninvasive Methods for Characterization of Large‐Volume Monolithic Chromatographic Columns

Podgornik, Aleš; Vidič, Jana; Jančar, Janez; Lendero, Nika; Frankovič, Vida; Štrancar, Aleš

doi:10.1002/ceat.200500170

Cited by 21 publications

(5 citation statements)

References 12 publications

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“…Since methacrylate monoliths of different pore sizes exhibit similar microstructure and porosity, differences in pressure drop reflect changes in monolith average pore size that determine the surface area and the DBC . Due to its noninvasivity and simplicity, pressure drop measurement can therefore be used as a fast and reliable method for testing the monolithic column properties . Pressure drop for 1 mL monolithic tubes of different pore sizes and functionalities as a function of flow rate is presented in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Although pressure drop–flow rate data can provide information on the size of monolith flow‐through pores and therefore represents simple and robust method for evaluating the performance of monolithic columns with different pore sizes, no information about functionalities and ligand densitiy is provided. pH transient response curve measurements can be a method of choice for such characterization as the method is noninvasive and simple . This method is based on the titration of ion‐exchange functional groups immobilized on the monolith pore surface with two buffers having the same pH value but different salt concentrations as commonly implemented in IEC.…”

Section: Resultsmentioning

confidence: 99%

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Effect of pore size on performance of monolithic tube chromatography of large biomolecules

et al. 2017

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Effect of pore size on the performance of ion-exchange monolith tube chromatography of large biomolecules was investigated. Radial flow 1 mL polymer based monolith tubes of different pore sizes (1.5, 2, and 6 μm) were tested with model samples such as 20 mer poly T-DNA, basic proteins, and acidic proteins (molecular weight 14 000-670 000). Pressure drop, pH transient, the number of binding site, dynamic binding capacity, and peak width were examined. Pressure drop-flow rate curves and dynamic binding capacity values were well correlated with the nominal pore size. While duration of the pH transient curves depends on the pore size, it was found that pH duration normalized on estimated surface area was constant, indicating that the ligand density is the same. This was also confirmed by the constant number of binding site values being independent of pore size. The peak width values were similar to those for axial flow monolith chromatography. These results showed that it is easy to scale up axial flow monolith chromatography to radial flow monolith tube chromatography by choosing the right pore size in terms of the pressure drop and capacity.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Effect of pore size on performance of monolithic tube chromatography of large biomolecules

et al. 2017

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“…Recently, several methods for nondestructive characterization were introduced [71,72]. While determination of pore size distribution and porosity can only be performed on a small monolith sample and can therefore be performed only during the production process adopting proper methodology [72], methods for determination of flow profile, ligand type, and ligand density can be per- formed by the end-user. Methods for determination of ligand type and density are based on the interaction between buffer ions and active group present on the monolith.…”

Section: Noninvasive Methods For Characterization Of Monolithic Columnsmentioning

confidence: 99%

Convective Interaction Media short monolithic columns: Enabling chromatographic supports for the separation and purification of large biomolecules

Barut¹,

Podgornik

Brne

et al. 2005

J of Separation Science

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New therapeutics that are being developed rely more and more on large and complex biomacromolecules like proteins, DNA, and viral particles. Manufacturing processes are being redesigned and optimized both upstream and downstream to cope with the ever-increasing demand for the above target molecules. In downstream processing, LC still represents the most powerful technique for achieving high yield and high purities of these molecules. In most cases, however, the separation technology relies on conventional particle-based technology, which has been optimized for the purification of smaller molecules. New technologies are, therefore, needed in order to push the downstream processing ahead and into the direction that will provide robust, productive, and easy to implement methods for the production of novel therapeutics. New technologies include the renaissance of membranes, various improvements of existing technologies, but also the introduction of a novel concept--the continuous bed or monolithic stationary phases. Among different introduced products, Convective Interaction Media short monolithic columns (SMC) that are based on methacrylate monoliths exhibit some interesting features that make them attractive for these tasks. SMC can be initially used for fast method development on the laboratory scale and subsequently efficiently transferred to preparative and even more importantly to industrial scale. A brief historical overview of methacrylate monoliths is presented, followed by a short presentation of theoretical considerations that had led to the development of SMC. The design of these columns, as well as their scale-up to large units, together with the methods for transferring gradient separations from one scale to another are addressed. Noninvasive methods that have been developed for the physical characterization of various batches of SMC, which fulfill the regulatory requirements for cGMP production, are discussed. The applications of SMC for the separation and purification of large biomolecules, which demonstrate the full potential of this novel technology for an efficient downstream processing of biomolecules, are also presented.

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“…However, polymethacrylate monoliths are the most utilized owing to their ease of synthesis, wide pH range, presence of epoxy ends, and mechanical and chemical stability . Numerous innovative studies into the advancement of monoliths through their formation processes, up‐scaling, non‐invasive characterization techniques, in situ analysis, and tackling of wall‐channelling have sprung up into the scene. These have maximized the usefulness of monoliths for bioscreening applications such as chromatographic separations, biosensors, catalytic bioreactors, purification, and concentration processes .…”

Section: Introductionmentioning

confidence: 99%

Thermogravimetric characterization of ex situ polymethacrylate (EDMA‐co‐GMA) monoliths

Acquah

Moy

Anwar³

et al. 2017

Can J Chem Eng

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The thermo-molecular mechanisms associated with free radical synthesis of polymethacrylate monoliths offer an effective pathway to tune their pore characteristics. In this work, thermogravimetric analyses were used for ex situ characterization of polymethacrylate monoliths synthesised from ethylene glycol dimethacrylate (EDMA) and glycidyl methacrylate (GMA). Apparent activation energies of the polymeric monoliths were determined by using Ozawa-Flynn-Wall (OFW) and Kissinger-Akahira-Sunose (KAS) isoconversional methods. The number of degradation stages for the polymers were observed to be as follows: homopolymeric GMA and EDMA (2 degradation stages each) and polymeric EDMA-co-GMA monoliths (3 degradation stages). The relationship between apparent activation energy and extent of conversion showed that the degradation of monoliths is based on a complex multi-step reaction rather than a single reaction model. Kinetic parameters showed that an increase in the composition of EDMA and GMA above 20 % significantly enhances the thermal stability of polymeric EDMA-co-GMA monoliths under elevated non-isothermal conditions. E40/G60 and E60/G40 monoliths were identified as the most thermally stable monoliths from the kinetic analyses.

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Noninvasive Methods for Characterization of Large‐Volume Monolithic Chromatographic Columns

Cited by 21 publications

References 12 publications

Effect of pore size on performance of monolithic tube chromatography of large biomolecules

Effect of pore size on performance of monolithic tube chromatography of large biomolecules

Convective Interaction Media short monolithic columns: Enabling chromatographic supports for the separation and purification of large biomolecules

Thermogravimetric characterization of ex situ polymethacrylate (EDMA‐co‐GMA) monoliths

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