New hydroxyapatite monolithic column for DNA extraction and its application in the purification of Bacillus subtilis crude lysate

Yu, Shengbing; Geng, Jianghui; Zhou, Ping; Wang, Jing; Chen, Xiangdong; Hu, Jiming

doi:10.1016/j.chroma.2007.11.120

Cited by 38 publications

(14 citation statements)

References 40 publications

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“…This further enhances their use as chromatographic adsorbent materials in the separation of viral and proteinaceous targets. Yu et al [149] constructed a miniaturised monolithic column with immobilised HA nanoparticles for rapid extraction of DNA molecules under optimised conditions of pH, ion type and concentration, and loading capacity. The optimised conditions and low salt requirement during the DNA extraction process enabled PCR amplification of PBE2 plasmid from Bacillus subtilis crude lysate.…”

Section: Hydroxyapatite Nanoparticles (Ha-np)mentioning

confidence: 99%

Nano-Doped Monolithic Materials for Molecular Separation

et al. 2017

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Monoliths are continuous adsorbents that can easily be synthesised to possess tuneable meso-/macropores, convective fluid transport, and a plethora of chemistries for ligand immobilisation. They are grouped into three main classes: organic, inorganic, and hybrid, based on their chemical composition. These classes may also be differentiated by their unique morphological and physicochemical properties which are significantly relevant to their specific separation applications. The potential applications of monoliths for molecular separation have created the need to enhance their characteristic properties including mechanical strength, electrical conductivity, and chemical and thermal stability. An effective approach towards monolith enhancement has been the doping and/or hybridization with miniaturized molecular species of desirable functionalities and characteristics. Nanoparticles are usually preferred as dopants due to their high solid phase dispersion features which are associated with improved intermolecular adsorptive interactions. Examples of such nanomaterials include, but are not limited to, carbon-based, silica-based, gold-based, and alumina nanoparticles. The incorporation of these nanoparticles into monoliths via in situ polymerisation and/or post-modification enhances surface adsorption for activation and ligand immobilisation. Herein, insights into the performance enhancement of monoliths as chromatographic supports by nanoparticles doping are presented. In addition, the potential and characteristics of less common nanoparticle materials such as hydroxyapatite, ceria, hafnia, and germania are discussed. The advantages and challenges of nanoparticle doping of monoliths are also discussed.

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Section: Hydroxyapatite Nanoparticles (Ha-np)mentioning

confidence: 99%

Nano-Doped Monolithic Materials for Molecular Separation

et al. 2017

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“…It is composed of functional groups of positively charged pairs of calcium ions and clusters of negatively charged phosphate groups. 90 It is essentially made up of the same materials that constitute bones and teeth, and so it is of "biosimilar" composition and is highly biocompatible. 91 The advantages of HA are that it is cheap, and biocompatible solid phases are available; cleaning in situ is feasible and it demonstrates good stability under alkaline conditions.…”

Section: 88mentioning

confidence: 99%

Technology advancements in antibody purification

Murphy¹,

Devine²,

O’Kennedy³

2016

ANTI

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Abstract:The separation of antibodies from complex mixtures can be achieved using chromatographic or non-chromatographic techniques. The purification of antibodies using chromatography involves the separation of antibodies or antibody-derived molecules present in complex mixtures by passing them through a solid phase (eg, silica resin or beads, monolithic columns, or cellulose membranes) and allowing the antibodies to bind or pass through depending on whether "bind-and-elute" or "flow-through" chromatographic methods are employed. Chromatographic methodologies can incorporate different separation techniques, such as affinity-tag binding, ion-exchange, size-exclusion chromatography, or immunoaffinity chromatography. However, in a concerted effort to become less reliant on chromatography-based separations owing to the high cost of large-scale production, non-chromatographic-based techniques such as precipitation, flocculation, crystallization, filtration, and aqueous two-phase partitioning are now becoming more popular. This review details current chromatographic and non-chromatographic methodo logies used for the purification of antibodies and expands on technological advancements and practical uses that have recently been reported.

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“…Table 1 compiles material data given by the manufacturer. Most recently, monolithic HA columns have been introduced [18]. HA is sensitive to acidic pH and prolonged exposure to pH below 6.5 will typically dissolve the material.…”

Section: The Materialsmentioning

confidence: 99%

“…Given the persistent manifestation of masstransfer limitations in the chromatography of pDNA, the monolithic HA column prepared by Yu et al [18] for DNA extraction from Bacillus subtilis crude lysates could present an interesting possibility. The monolith skeleton had micrometer pores and was prepared by a sol-gel technique.…”

Section: Plasmid Dna Purificationmentioning

confidence: 99%

Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography

Hilbrig

Freitag

2011

Biotechnology Journal

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Hydroxyapatite and related stationary phases increasingly play a role in the downstream processing of high-value biological materials, such as recombinant proteins, therapeutic antibodies and pharmaceutical-grade plasmid DNA. Chromatographic hydroxyapatite is an inorganic, ceramic material identical in composition, if not in structure, to calcium phosphate found in human bones and teeth. The interaction of hydroxyapatite with biomacromolecules is complex and highly dynamic, which can make predicting performance difficult, but also allows the design of very selective isolation processes. This review discusses the currently commercially available chromatographic materials, different retention mechanisms supported by these materials and differential exploitation for the design of highly specific isolation procedures. The state of the art of antibody purification by hydroxy-and fluoroapatite is reviewed together with tested routines for method development and implementation. Finally, the isolation of plasmid DNA is discussed, since the purification of DNA therapeutics at a sufficiently large scale is an emerging need in bioprocess development and perhaps the area in bioseparation where apatite chromatography can make its most important contribution to date.

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New hydroxyapatite monolithic column for DNA extraction and its application in the purification of Bacillus subtilis crude lysate

Cited by 38 publications

References 40 publications

Nano-Doped Monolithic Materials for Molecular Separation

Nano-Doped Monolithic Materials for Molecular Separation

Technology advancements in antibody purification

Isolation and purification of recombinant proteins, antibodies and plasmid DNA with hydroxyapatite chromatography

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