DNA purification by triple‐helix affinity precipitation

Costioli, Matteo; Fisch, Igor; Garret‐Flaudy, Frédéric; Hilbrig, Frank; Freitag, Ruth

doi:10.1002/bit.10497

Cited by 87 publications

(81 citation statements)

References 43 publications

(48 reference statements)

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“…The formation of a triple helix by an immobilized oligonucleotide and a DNA double strand were used for the purification of nucleic acids by Ito et al [56]. Schluep and Cooney [57,58] as well as Costioli et al [59] have used it for selective purification of plasmid DNA. However, the method is suffering from the slow formation of the triple helix and the low chemical and biochemical stability of the immobilized oligonucleotide.…”

Section: Affinity Separationmentioning

confidence: 99%

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

Voss

2008

Chem Eng & Technol

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Plasmid DNA is used as a cloning vector to deliver recombinant genetic information into microorganisms. Since the 1990s, this principle has also been applied for the delivery of therapeutic genes in gene therapy and genetic vaccination. This non-viral gene delivery is afflicted with fewer safety concerns in comparison to viral systems. Processes for the production of high-quality plasmid DNA at multiand kilogram scale are necessary to meet the needs of clinical trials as well as future therapeutics. Cell disruption, the separation of structurally-related impurities and analytical techniques for process and quality control are the main challenges for bioengineering. This review summarizes the development in these fields over the past recent years.

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Section: Affinity Separationmentioning

confidence: 99%

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

Voss

2008

Chem Eng & Technol

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“…Costioli and co-workers recently presented an affinity precipitation method based on triplehelix formation (Costioli et al, 2003). An affinity ligand was coupled to a stimulus-responsive polymer that on a small change in the temperature changes its solubility (Costioli et al, 2003). Although these new selective precipitation techniques have been reported, none is working directly on a clarified lysate prepared by alkaline lysis in the presence of the high concentrations of potassium acetate.…”

Section: Introductionmentioning

confidence: 98%

“…In another method a cationic detergent, CTAB, was used to selectively precipitate plasmid DNA from RNA, proteins, and endotoxins (Lander et al, 2002). Costioli and co-workers recently presented an affinity precipitation method based on triplehelix formation (Costioli et al, 2003). An affinity ligand was coupled to a stimulus-responsive polymer that on a small change in the temperature changes its solubility (Costioli et al, 2003).…”

Section: Introductionmentioning

confidence: 98%

Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate

Wahlund¹,

Gustavsson

Izumrudov³

et al. 2004

Biotech & Bioengineering

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The demand for highly purified plasmids in gene therapy and plasmid-based vaccines requires large-scale production of pharmaceutical-grade plasmid. Large-scale purification of plasmid DNA from bacterial cell culture normally includes one or several chromatographic steps. Prechromatographic steps include precipitation with solvents, salts, and polymers combined with enzymatic degradation of nucleic acids. No method alone has so far been able to selectively capture plasmid DNA directly from a clarified alkaline lysate. We present a method for selective precipitation of plasmid DNA from a clarified alkaline lysate using polycation poly(N, N'-dimethyldiallylammonium) chloride (PDMDAAC). The specific interaction between the polycation and the plasmid DNA resulted in the formation of a stoichiometric insoluble complex. Efficient removal of contaminants such as RNA, by far the major contaminant in a clarified lysate, and proteins as well as 20-fold plasmid concentration has been obtained with about 80% recovery. The method utilizes a inexpensive, commercially available polymer and thus provides a capture step suitable for large-scale production.

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“…11,12 Since then, a number of applications have been realized with this class of materials in addition to gene or ODN delivery. These range from purification of biomaterials 13 to DNA detection. 14 The appropriate selection of organic polymer and DNA sequence allows tuning and controlling of the material properties for designated applications.…”

Section: Introductionmentioning

confidence: 99%

DNA Block Copolymers: Functional Materials for Nanoscience and Biomedicine

2012

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W e live in a world full of synthetic materials, and the development of new technologies builds on the design and synthesis of new chemical structures, such as polymers. Synthetic macromolecules have changed the world and currently play a major role in all aspects of daily life. Due to their tailorable properties, these materials have fueled the invention of new techniques and goods, from the yogurt cup to the car seat belts. To fulfill the requirements of modern life, polymers and their composites have become increasingly complex. One strategy for altering polymer properties is to combine different polymer segments within one polymer, known as block copolymers. The microphase separation of the individual polymer components and the resulting formation of well defined nanosized domains provide a broad range of new materials with various properties. Block copolymers facilitated the development of innovative concepts in the fields of drug delivery, nanomedicine, organic electronics, and nanoscience.Block copolymers consist exclusively of organic polymers, but researchers are increasingly interested in materials that combine synthetic materials and biomacromolecules. Although many researchers have explored the combination of proteins with organic polymers, far fewer investigations have explored nucleic acid/polymer hybrids, known as DNA block copolymers (DBCs). DNA as a polymer block provides several advantages over other biopolymers. The availability of automated synthesis offers DNA segments with nucleotide precision, which facilitates the fabrication of hybrid materials with monodisperse biopolymer blocks. The directed functionalization of modified single-stranded DNA by WatsonÀCrick base-pairing is another key feature of DNA block copolymers. Furthermore, the appropriate selection of DNA sequence and organic polymer gives control over the material properties and their self-assembly into supramolecular structures. The introduction of a hydrophobic polymer into DBCs in aqueous solution leads to amphiphilic micellar structures with a hydrophobic polymer core and a DNA corona.In this Account, we discuss selected examples of recent developments in the synthesis, structure manipulation and applications of DBCs. We present achievements in synthesis of DBCs and their amplification based on molecular biology techniques. We also focus on concepts involving supramolecular assemblies and the change of morphological properties by mild stimuli. Finally, we discuss future applications of DBCs. DBC micelles have served as drug-delivery vehicles, as scaffolds for chemical reactions, and as templates for the self-assembly of virus capsids. In nanoelectronics, DNA polymer hybrids can facilitate size selection and directed deposition of single-walled carbon nanotubes in field effect transistor (FET) devices.

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DNA purification by triple‐helix affinity precipitation

Cited by 87 publications

References 43 publications

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

Downstream Processing of Plasmid DNA for Gene Therapy and Genetic Vaccination

Precipitation by polycation as capture step in purification of plasmid DNA from a clarified lysate

DNA Block Copolymers: Functional Materials for Nanoscience and Biomedicine

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