Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography

Belen'kii, B.G.; Podkladenko, A.M.; Kurenbin, O.I.; Mal’tsev, V. G.; Nasledov, D. G.; Trushin, Sergei A.

doi:10.1016/0021-9673(93)80613-d

Cited by 44 publications

(28 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A first consequence was a change in the phraseology, for example, a change from the term "column length" to the concept of the "operative thickness (length) of adsorption layer", which corresponds to the minimum distance necessary to achieve a given separation. A detailed analysis of the behavior of proteins in gradient elution HPMDC can be found in some recent publications [e. g. 19,31,37,38]. It was found that the behavior of proteins in gradient elution HPMDC could in principle be treated by the same set of chromatographic theorems as conventional gradient HPLC, which incidentally demonstrates again that there is no basic difference between HPMDC and liquid chromatography in general.…”

Section: Hpmdc Of Proteins: a Single-step Desorption Processmentioning

confidence: 95%

An Introduction to Monolithic Disks as Stationary Phases for High Performance Biochromatography

Tennikova

Freitag

2000

J. High Resol. Chromatogr.

View full text Add to dashboard Cite

Section: Hpmdc Of Proteins: a Single-step Desorption Processmentioning

confidence: 95%

An Introduction to Monolithic Disks as Stationary Phases for High Performance Biochromatography

Tennikova

Freitag

2000

J. High Resol. Chromatogr.

View full text Add to dashboard Cite

“…Modified poly(glycidyl methacrylate-ethylene dimethacrylate) monoliths have been applied in form of small disks for separation of proteins. The concept of a short layer is based on the fact that after a certain distance in the column, the zone migrates with the same velocity as the mobile phase and does not further contribute to resolution [26,27]. High resolution can be obtained in such a short column.…”

Section: Introductionmentioning

confidence: 98%

Mass transfer characteristics of plasmids in monoliths

Zöchling

Hahn

Ahrer

et al. 2004

J of Separation Science

View full text Add to dashboard Cite

The hydrodynamic properties and pore-structure of monoliths based on functionalized poly(glycidyl methacrylate-ethylene dimethacrylate) were characterised by pulse response experiments using different probes representing a wide range of molecular mass. On a small scale, band spreading was found to be caused to the extent of more than 90% by extra-column effects. These monoliths have large channel diameters, providing a suitable chromatography adsorbent for processing of large molecules. Dynamic and static binding capacity for plasmid DNA was investigated. For our model plasmid, consisting of 4.9 kbp, a capacity of 7 mg/mL was observed in comparison to 0.3 mg/mL for a conventional medium designed for protein separation. When plasmids were loaded on the monolith a gradual increase in pressure drop was observed. The channels filled up and the cross-sectional area available for liquid flow decreased. Therefore, a higher pressure drop was observed during elution. This is caused by (i) shrinking of the channels as effect of the high salt concentration, (ii) high viscosity of the mobile phase due to high concentration of plasmids, and (iii) an increase of the hydrodynamic radius of the plasmid with salt concentration from 45 nm at 150 mM to 70 nm at 2 M NaCl, as measured by dynamic light scattering. These types of monoliths are considered to be the preferred adsorbents for plasmid separation.

show abstract

“…These results were further elaborated by Belenkii et al [44], who presented an approach for the estimation of the critical length of a column in gradient HPLC of proteins. Their approach was based on the concept of critical chromatography of synthetic polymers.…”

Section: Historical Overviewmentioning

confidence: 63%

“…The results obtained by Tennikova and coworkers [45] for ion-exchange chromatography and by Belenkii et al [44] for RP chromatography form a theoretical basis for the chromatography on SMC. The main conclusion from these studies was that it is possible to use ultra-short (in a millimeter range) columns for an efficient separation of proteins and other molecules with appropriate resolution and short gradient times.…”

Section: Historical Overviewmentioning

confidence: 83%

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

View full text Add to dashboard Cite

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.

show abstract

Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography

Cited by 44 publications

References 32 publications

An Introduction to Monolithic Disks as Stationary Phases for High Performance Biochromatography

An Introduction to Monolithic Disks as Stationary Phases for High Performance Biochromatography

Mass transfer characteristics of plasmids in monoliths

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

Contact Info

Product

Resources

About