Journal ArticleDOI
Methacrylate‐based short monolithic columns: Enabling tools for rapid and efficient analyses of biomolecules and nanoparticles
Miloš Barut,Aleš Podgornik,Lidija Urbas,Boštjan Gabor,Peter Brne,Jana Vidič,Sašo Plevčak,Aleš Štrancar +7 more
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TLDR
The efficiency of the monolithic column is demonstrated--where the mass transfer between the stationary and mobile phase is greatly enhanced--for the in-process and final control of the new therapeutics.Abstract:
This review describes the novel chromatography stationary phase - a porous monolithic methacrylate-based polymer - in terms of the design of the columns and some of the features that make these columns attractive for the purification of large biomolecules. We first start with a brief summary of the characteristics of these large molecules (more precisely large proteins like immunoglobulins G and M, plasmid deoxyribonucleic acid (DNA), and viral particles), and a list of some of the problems that were encountered during the development of efficient purification processes. We then briefly describe the structure of the methacrylate-based monolith and emphasize the features which make them more than suitable for dealing with large entities. The highly efficient structure on a small scale can be transferred to a large scale without the need of making column modifications, and the various approaches of how this is accomplished are briefly presented in this paper. This is followed by presenting some of the examples from the bioprocess development schemes, where the implementation of the methacrylate-based monolithic columns has resulted in a very efficient and productive process. Following this, we move back to the analytical scale and demonstrate the efficiency of the monolithic column - where the mass transfer between the stationary and mobile phase is greatly enhanced - for the in-process and final control of the new therapeutics. The combination of an efficient structure and the appropriate hardware results in separations of proteins with residence time less than 0.1 s.read more
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Journal ArticleDOI
High Throughput Isolation and Glycosylation Analysis of IgG–Variability and Heritability of the IgG Glycome in Three Isolated Human Populations
Maja Pučić,Ana Knezevic,Jana Vidič,Barbara Adamczyk,Mislav Novokmet,Ozren Polasek,Olga Gornik,Sandra Supraha-Goreta,Mark R. Wormald,Irma Redzic,Harry Campbell,Alan F. Wright,Nicholas D. Hastie,James F. Wilson,Igor Rudan,Manfred Wuhrer,Pauline M. Rudd,Djuro Josic,Djuro Josic,Gordan Lauc +19 more
TL;DR: The majority of the structural features of the IgG glycome were consistent with previous studies, but sialylation was somewhat higher than reported previously, indicating that the final glycan structures are not a simple result of competing enzymatic activities, but a carefully regulated outcome designed to meet the prevailing physiological needs.
Journal ArticleDOI
Applications of polymethacrylate-based monoliths in high-performance liquid chromatography.
E. G. Vlakh,Tatiana B. Tennikova +1 more
TL;DR: In this review, the applications of polymethacrylate-based monolithic columns are summarized for the separation, purification and analysis of low and high molecular mass compounds in the different HPLC formats, including micro- and large-scale HPLC modes.
Journal ArticleDOI
Current trends in separation of plasmid DNA vaccines: A review
TL;DR: This review summarizes the current trends in the purification of pDNA vaccines for practical and analytical applications and special attention is paid to chromatographic techniques aimed at reducing the steps of final purification, post primary isolation and intermediate recovery.
Journal ArticleDOI
Fast separation of large biomolecules using short monolithic columns.
TL;DR: The most efficient separation is expected when short monolithic column with accordingly adjusted gradient is implemented, especially for macromolecules interacting with the monolith functionalities via over 10 binding sites.
Journal ArticleDOI
Advances in chromatographic supports for pharmaceutical‐grade plasmid DNA purification
TL;DR: The present review discusses the structural progress and evolution of the chromatographic supports that have been used for plasmid DNA purification, and focuses on the chemical and structural classification of the different media and on some of the specific ligands used forplasmidDNA bioseparation.
References
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Continuous rods of macroporous polymer as high-performance liquid chromatography separation media
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Octadecylsilylated Porous Silica Rods as Separation Media for Reversed-Phase Liquid Chromatography
TL;DR: The performance of the silica rods was much better at a high flow rate than that of conventional columns packed with 5 μm C(18) silica particles having 12 and 30 nm pores, especially for high molecular weight species.
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New designs of macroporous polymers and supports : From separation to biocatalysis
TL;DR: Polymer monoliths containing intricate pore networks can be obtained in any desired shape by a simple molding process and provide unique advantages such as fast kinetics, high reactions, and high throughput.
Journal ArticleDOI
Monolithic, “Molded”, Porous Materials with High Flow Characteristics for Separations, Catalysis, or Solid-Phase Chemistry: Control of Porous Properties during Polymerization
TL;DR: The porosity and flow characteristics of macroporous polymer monoliths that may be used to prepare separation media, flow-through reactors, catalysts, or supports for solid-phase chemistry can be controlled easily during their preparation.
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Applications of polymethacrylate-based monoliths in high-performance liquid chromatography.
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