Improved purification of recombinant adenoviral vector by metal affinity membrane chromatography

Lee, Dong-Sop; Kim, Byong-Moon; Seol, Dai-Wu

doi:10.1016/j.bbrc.2008.11.096

Cited by 29 publications

(18 citation statements)

References 8 publications

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“…Zinc employed as an ion‐pairing agent forms a complex coordination structure with oleate to produce a solid matrix, where 1 zinc cation interacts with 4 oleate anions (34). As a cation, zinc also has exceptionally high affinity for Ad previously exploited for vector purification (35). Its essential role in supporting the high transduction capacity of MNP(Ad)s is evidenced by the substantially greater gene transfer efficiency of the latter in comparison to particles prepared with calcium instead of zinc as the ion‐pairing agent (See Supplemental Fig.…”

Section: Discussionmentioning

confidence: 99%

Site‐specific gene delivery to stented arteries using magnetically guided zinc oleate‐based nanoparticles loaded with adenoviral vectors

et al. 2013

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Gene therapeutic strategies have shown promise in treating vascular disease. However, their translation into clinical use requires pharmaceutical carriers enabling effective, site-specific delivery as well as providing sustained transgene expression in blood vessels. While replication-deficient adenovirus (Ad) offers several important advantages as a vector for vascular gene therapy, its clinical applicability is limited by rapid inactivation, suboptimal transduction efficiency in vascular cells, and serious systemic adverse effects. We hypothesized that novel zinc oleate-based magnetic nanoparticles (MNPs) loaded with Ad would enable effective arterial cell transduction by shifting vector processing to an alternative pathway, protect Ad from inactivation by neutralizing factors, and allow site-specific gene transfer to arteries treated with stent angioplasty using a 2-source magnetic guidance strategy. Ad-loaded MNPs effectively transduced cultured endothelial and smooth muscle cells under magnetic conditions compared to controls and retained capacity for gene transfer after exposure to neutralizing antibodies and lithium iodide, a lytic agent causing disruption of free Ad. Localized arterial gene expression significantly stronger than in control animal groups was demonstrated after magnetically guided MNP delivery in a rat stenting model 2 and 9 d post-treatment, confirming feasibility of using Ad-loaded MNPs to achieve site-specific transduction in stented blood vessels. In conclusion, Ad-loaded MNPs formed by controlled precipitation of zinc oleate represent a novel delivery system, well-suited for efficient, magnetically targeted vascular gene transfer.

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Section: Discussionmentioning

confidence: 99%

Site‐specific gene delivery to stented arteries using magnetically guided zinc oleate‐based nanoparticles loaded with adenoviral vectors

et al. 2013

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“…These include detergent cell lysis (Peixoto et al, 2006), DNA digestion (Shabram et al, 1997), depth filter clarification (Goerke et al, 2005;Peixoto et al, 2008), Ad capture by anion exchange (Duffy et al, 2005;Peixoto et al, 2008) and Ad Characterization of a representative lot of Ad using the assays described in Materials and Methods Section. The first step is typically an anion exchange resin or membrane (Duffy et al, 2005;Peixoto et al, 2008) and the second a gel filtration (Eglon et al, 2009), ionexchange (Blanche et al, 2000) metal chelate resin (Huyghe et al, 1995), or metal chelate membrane (Lee et al, 2009). Two column processes are described in the literature.…”

Section: Discussionmentioning

confidence: 99%

Development of a platform process for adenovirus purification that removes human set and nucleolin and provides high purity vector for gene delivery

Riske

Berard²,

Albee³

et al. 2012

Biotech & Bioengineering

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The manufacturing of virus occurs at a modest scale in comparison to many therapeutic proteins mainly because a gene therapy dose is typically only µg of vector. Although modest in scale the generation of high purity virus is challenging due to low viral expression levels and the difficulties in adequately characterizing such a large and complex molecule. A 100 L bioreactor might produce only 100 mg of virus that must be separated from host and process impurities that are typically greater by several orders of magnitude. Furthermore, in the later purification stages the main milieu component is often virus at low concentration (µg/mL) which may non-specifically adsorb to purification surfaces resulting in a lowered virus recovery. This study describes our approach to develop a scalable, manufacturable robust process for an Adenovirus (Ad) gene therapy vector. A number of analytical tools were developed to guide the purification design. During process development, two human proteins, SET and nucleolin, were identified in viral preparations. To our knowledge, this is the first time that SET and nucleolin have been described in Ad. In this report we detail a process for their removal and the robust removal of all process, product and host cell impurities.

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“…However, an additional AEC step was necessary to complete DNA removal, yielding about 1.4 µg DNA/mg VLP in the final product. Sart IMAC was found insufficient to reduce the level of DNA contaminants co-eluting with AdV particles in packed-bed AEC [174]. Treatment of the AdV pool with Benzonase before the second chromatographic step remained necessary to lower significantly the DNA concentration in the final MA pool.…”

Section: Sartobind® Iec/afc Masmentioning

confidence: 95%

“…In agreement with the above trends, some general results from these works can be underlined. MAs gave improved virus yields and/or productivities compared to conventional packed beads [39,43,166,169,174]. Strong membrane ion exchangers (Q, S) were more efficient than weak ones (D, C) [43,163,167].…”

Section: Sartobind® Iec/afc Masmentioning

confidence: 99%

“…Residual DNA impurities were not eliminated, however [164,176]. The amount of contaminating DNA could be lowered by nuclease pretreatment, which was applied in some studies [165,174,177] and suggested in others [39,43,166,176], and/or by the combination of chromatographic steps. Thus, Vicente et al [175] used SEC with Sephacryl™ S-500 resin to polish the pool of rotavirus-like particles captured by Sart D AEC.…”

Section: Sartobind® Iec/afc Masmentioning

confidence: 99%

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Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Junter

Lebrun

2020

Journal of Pharmaceutical Analysis

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Improved purification of recombinant adenoviral vector by metal affinity membrane chromatography

Cited by 29 publications

References 8 publications

Site‐specific gene delivery to stented arteries using magnetically guided zinc oleate‐based nanoparticles loaded with adenoviral vectors

Site‐specific gene delivery to stented arteries using magnetically guided zinc oleate‐based nanoparticles loaded with adenoviral vectors

Development of a platform process for adenovirus purification that removes human set and nucleolin and provides high purity vector for gene delivery

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

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