Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity

Yu, Mengran; Li, Yan; Zhang, Songping; Li, Xiunan; Yang, Yanli; Chen, Yi; Ma, Guanghui; Su, Zhiguo

doi:10.1016/j.chroma.2014.01.027

Cited by 42 publications

(15 citation statements)

References 51 publications

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“…Biomanufacturing of the viral vectors at present suffers from low process yields due 29 to suboptimal chromatography processes due to inaccessibility of the resin pores for large solutes 30 such as viral vectors resulting in an order of magnitude lower binding capacities compared to proteins 31 [1]. New materials with a more open porous architecture such as gigaporous resins [2], monoliths [3, of the retentate with PBS followed 2-fold concentration thus each diafiltration step corresponding to 148 one diavolume of the buffer exchanged. A transmembrane pressure of 0.6 bars and the maximum 149 stirring speed of 1500 rpm was used for the initial concentration step and subsequent diafiltration 150 steps.…”

Section: Introduction 27mentioning

confidence: 99%

Characterisation of porous anodic alumina membranes for ultrafiltration of protein nanoparticles as a size mimic of virus particles

Sharma

Bracewell

2019

Journal of Membrane Science

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9Viral vectors used in emerging gene therapies face challenges of significant yield loss during 10 downstream processing primarily due to their large size, fragility and mass transfer limitations of the 11 traditional porous chromatography adsorbents. The large size of the vectors in relation to key 12 impurities makes them suitable solutes for ultrafiltration-based separations. Efforts to utilise 13 ultrafiltration for virus purification are often restricted to commercial polymeric membranes with wide 14 pore size distributions and tortuous, interconnected channels. Membranes with narrow pore 15 distributions and straight pore channels such as porous anodic alumina (PAA) may present 16 opportunities for improved virus purification. This paper examines the use of porous anodic alumina 17 membranes for application in virus separation by using model solutes such as thyroglobulin and 18 protein nanoparticles. A systematic approach is used to select a polymeric ultrafiltration membrane 19 rating for comparison with 20nm rated PAA membrane by comparing hydraulic permeability and 20 dextran sieving characteristics of the membranes. Differences in the filterability of the model solutes 21 were characterised. Finally, a discontinuous diafiltration experiment was employed to fractionate 22 smaller model impurity and protein nanoparticles. Results indicate that PAA membranes have 23 superior fouling resistance with 3-4 folds higher flux recovery ratio and 3-fold higher purification 24 factors compared to the polymeric membranes, but the presence of surface defects make them more 25 susceptible to product loss through leaky transmission. 26 explore the use of ultrafiltration based processing instead of chromatography based processes by exploiting differences in the size of impurities and viral particles [6][7][8][9]. Most of the reported literature 35 compares different molecular weight rating or pore size ratings of polymeric ultrafiltration membranes 36 or different modules such as hollow fibre and cassette [6, 8]. These membranes have wide pore size 37 distribution, which is often reported to affect the membrane performance especially retention of large 38 biomolecules such as viruses [10, 11]. Isoporous membranes with different porous architectures such 39 as porous anodic alumina (PAA) membranes are however being investigated in other fields such as 40 nanofiltration for water purification and biosensors. 41Porous anodic alumina membranes are widely studied for applications in label-free biosensors [12][13][14] 42 for use as point-of-care diagnostic devices. Applications of PAA membranes in bioseparations have 43 been limited to diffusion based separations for haemodialysis [15], separation of similarly sized 44 proteins by exploiting differences in solute charge [16], enriching phosphoproteins for mass 45 spectrometry [17] and ultrafiltration of small proteins [18-20]. A few reports on PAA membranes have 46 examined their potential for virus separations. Moon et al., [21] reported the ability of 35-nm pores of a 47 PA...

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Section: Introduction 27mentioning

confidence: 99%

Characterisation of porous anodic alumina membranes for ultrafiltration of protein nanoparticles as a size mimic of virus particles

Sharma

Bracewell

2019

Journal of Membrane Science

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“…Tangential flow filtration (TFF) methods have been used to concentrate and purify influenza virus, a retrovirus vector, and AAV using size exclusion as the main mechanism of separation with recoveries up to 100%. Adsorption mechanisms, ranging from chromatography beads to monoliths are also often applied to viral particles. The recovery from adsorption methods is usually lower than filtration methods, with a typical range between 30 and 68%.…”

Section: Introductionmentioning

confidence: 99%

A generalized purification step for viral particles using mannitol flocculation

Heldt

Saksule

Joshi

et al. 2018

Biotechnology Progress

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Vaccine manufacturing has conventionally been performed by the developed world using traditional unit operations like filtration and chromatography. There is currently a shift in the manufacturing of vaccines to the less developed world, requiring unit operations that reduce costs, increase recovery, and are amenable to continuous manufacturing. This work demonstrates that mannitol can be used as a flocculant for an enveloped and nonenveloped virus and can purify the virus from protein contaminants after microfiltration. The recovery of the virus ranges from 58 to 96% depending on virus, the filter pore size, and the starting concentration of the virus. Protein removal of 80% was achieved for the small nonenveloped virus using a 0.1 µm filter because proteins were not flocculated with the virus and flowed through the filter. It is hypothesized that mannitol dehydrates the viral surface by controlling the water structure surrounding the virus. Without the ability to become compact, as occurs with proteins, the virus aggregates in the presence of osmolytes and proteins do not. Osmolyte flocculation is a scalable process using high flux microfilters. It has been applied to both an enveloped and nonenveloped virus, making this process friendly to a variety of vaccine and gene therapy products. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1027-1035, 2018.

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“…By contrast, the microscopic methods are based on the determination of intraparticle concentration profiles, and thus could provide more insight into the transport mechanism in a particle scale . In the past two decades, microscopic researches were widely performed on individual ion‐exchange particles by using confocal laser scanning microscopy and optical microscopy to visualize the transient patterns of protein adsorption in particles and protein transfer rate could be determined based on the visualizations. Up to now, the vast majority of microscopic measurements have been usually restricted to optically transparent spherical particles.…”

Section: Introductionmentioning

confidence: 99%

Visualization and Modeling of Protein Adsorption and Transport in DEAE‐ and DEAE‐Dextran‐Modified Bare Capillaries

Xue

Sun

2018

AIChE Journal

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This article reports a microscopic method to directly visualize protein adsorption and transport in bare capillaries modified with ion exchange groups. Silica capillaries were coated with poly(vinyl alcohol) to eliminate nonspecific protein absorption and then functionalized with diethylaminoethyl (DEAE) and DEAE-dextran to prepare anion-exchange capillaries denoted as D-C and D-dex-C, respectively. Protein concentration profiles acquired from microscopic images were analyzed with a one-dimensional diffusion-adsorption model to determine adsorption capacities and effective diffusivities. It revealed that surface diffusion of bound protein on the capillary surface in D-Cs was negligible, but that of bound proteins markedly contributed to protein transfer in D-dex-Cs because of the chain delivery effect of charged dextran chains. The chain delivery was little affected by protein concentration, but significantly increased with ionic capacity. The research thus provided new insight into protein adsorption and transport behaviors on the surfaces modified with ion exchange groups of different structures.

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Improving stability of virus-like particles by ion-exchange chromatographic supports with large pore size: Advantages of gigaporous media beyond enhanced binding capacity

Cited by 42 publications

References 51 publications

Characterisation of porous anodic alumina membranes for ultrafiltration of protein nanoparticles as a size mimic of virus particles

Characterisation of porous anodic alumina membranes for ultrafiltration of protein nanoparticles as a size mimic of virus particles

A generalized purification step for viral particles using mannitol flocculation

Visualization and Modeling of Protein Adsorption and Transport in DEAE‐ and DEAE‐Dextran‐Modified Bare Capillaries

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