Author
Peter Brne
Bio: Peter Brne is an academic researcher from BIA Separations (Slovenia). The author has contributed to research in topics: Monolithic HPLC column & Ion chromatography. The author has an hindex of 8, co-authored 8 publications receiving 330 citations.
Papers
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TL;DR: Convective Interaction Media short monolithic columns (SMC) that are based on methacrylate monoliths exhibit some interesting features that make them attractive for these tasks, and applications of SMC for the separation and purification of large biomolecules are presented.
Abstract: 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.
71 citations
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TL;DR: 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.
58 citations
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TL;DR: It was concluded that for a given monolith the protein capacity can be derived from the data obtained by the new method, which was shown to be suitable for determining the amount of ionic groups on both anion and cation monolithic columns.
53 citations
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TL;DR: The duration of pH transient was shown to be linearly proportional to the total ionic capacity and was used to estimate protein dynamic binding capacity of the resin and to distinguish between cation and anion or weak and strong ion exchange chromatographic supports.
43 citations
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TL;DR: The optimization of conditions for selective depletion of HSA and IgG using affinity and pseudo-affinity chromatography and two-dimensional gel electrophoresis is described.
38 citations
Cited by
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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.
422 citations
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TL;DR: This review discusses the column efficiency of HILIC materials in relation to solute and stationary phase structures, as well as comparisons between particle-packed and monolithic columns.
394 citations
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TL;DR: Technology trends in antibody purification are reviewed, including recent innovations in size exclusion, anion exchange, cation exchange, hydrophobic interaction, immobilized metal affinity, mixed-mode, and bioaffinity chromatography.
264 citations
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TL;DR: The pressure drop characteristics, mass transfer properties, scale-up, and applications of monoliths in the context of conventional chromatography media are discussed.
218 citations
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TL;DR: In this work, some aspects of the development of mixed-mode chromatography are reviewed, such as stationary phase preparation, combinations of various separation modes, separation mechanisms, typical applications to biopolymers and peptides, and future prospects.
155 citations