An efficient capture strategy for the purification of human adenovirus type 5 from cell lysates

Dietl, Sandra; Kiefer, Franziska; Binder, Stefanie Petrou; Walther, Paul; Sobek, Harald; Mizaikoff, Boris

doi:10.1016/j.jbiotec.2022.11.015

Cited by 6 publications

(2 citation statements)

References 34 publications

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“…The total DNA and protein reduction were remarkable in both cases, indicating robustness of the process toward final‐product purity. Moreover, the concentration of the virus was comparable if not slightly better than that reported in the literature, and in both cases, the recovery was higher, and the removal of DNA and proteins was more efficient [25].…”

Section: Resultsmentioning

confidence: 92%

Bridging upstream and downstream for improved adenovirus 5 bioprocess

Petrović Koshmak,

Jug,

Vrabec

et al. 2023

Electrophoresis

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Adenoviruses are well‐known viral vectors that have been previously used in gene therapy and as a vaccine‐delivery vehicle for humans and animals. During the COVID‐19 pandemic, it gained renewed attention, but at the same time, it raised concerns due to side effects observed with some of the resulting vaccines administered to patients. It has been indicated that these side effects might be attributed to impurities present in the final product. Therefore, constant enhancement of the vaccine purity and further improvement of impurity detection methods are needed. In this work, we showcase an example of industry‐relevant adenovirus bioprocess optimization. Our data show the effect of upstream parameters on the bioburden introduced to the downstream process. We provide an example of process optimization using a combination of the PATfix analytical method, ddPCR, infectivity, total DNA, and total protein analyses to optimize cell density, multiplicity of infection, and length of production. Additionally, we provide data illustrating the robustness of the convective interaction media quaternary amine monolithic chromatography step. This anion exchange strategy was shown to remove over 99% of protein and DNA impurities, including those unable to be addressed by tangential flow filtration, while maintaining high adenovirus recoveries.

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Section: Resultsmentioning

confidence: 92%

Bridging upstream and downstream for improved adenovirus 5 bioprocess

Petrović Koshmak,

Jug,

Vrabec

et al. 2023

Electrophoresis

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“…As a widely used equipment for deep gas-liquid separation, the coalescer is utilized in various fields such as chemical [1], medicine [2], and petroleum [3]. Researchers have conducted extensive studies on the structural parameters that affect its separation performance, including pore size gradient, thickness, fiber diameter, and filler density.…”

Section: Introductionmentioning

confidence: 99%

Experimental study on the optimum number of layers of fiber mats for gas-liquid coalescer

Chaolei,

Zhiqian,

Zhenbo

et al. 2024

J. Phys.: Conf. Ser.

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The coalescer is widely used in deep gas dehydration due to its high efficiency. Most manufacturers make the filter by enwinding multiple layers of fiber mats to obtain high efficiency, which leads to high-pressure losses in practical applications. Obtaining the number of layers of fiber mats is based on engineering practice and experiences, which needs more data support and theoretical basis. 1-10 layers of hydrophilic fiber mat samples were made, and experiments were carried out at filter face velocities of 0.14m/s to 0.35m/s and liquid loading rate of 1.25g/min to 5g/min. In addition, the concept of collision probability based on fiber porosity is invoked. A model between efficiency and porosity is developed to obtain the optimal solution for the number of layers. The results show that the saturation is monotonically decreasing, while the pressure drop linearly increases as velocity increases. The optimum number of fiber mat layers decreases from 5 to 3 as the filter face velocity increases. The efficiency model based on the fiber porosity agrees with the experimental efficiency values, which provides theoretical support for calculating the optimal number of fiber mat layers.

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3D‐Printed Highly Porous Functional Materials for the Efficient Removal of Adenovirus

Keitel,

Dietl,

Philipp

et al. 2024

Adv Materials Technologies

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Hierarchical porous acrylate‐based materials are highly interesting as 3D filter materials, such as for virus removal from suspensions. Here, the synthesis of highly porous monolithic 3D materials by polymerization‐induced phase separation in liquid crystal display (LCD) based 3D printing is presented for the efficient removal of human adenovirus type 5. The hierarchical porosity can be tuned via the variation of the photocurable resin composition (i.e., inherent porosity) and the computer‐aided design (i.e., “printed” porosity; microchannels). 3D polymer structures with highly intricate geometries and structural features ranging from ≈20 nm up to cm can be achieved, which can be used for effective virus removal in a laboratory‐scale flow‐through approach. Combined focused ion beam/scanning electron microscopy tomography and mercury porosimetry provide detailed information on the inherent pore size, pore size distribution, and pore interconnectivity, which is key for the performance of such functional 3D materials. Polymers with a theoretical void volume of 75% show virus capture with a removal efficiency of ≈70% of the adenovirus. Polymers with the same theoretical void volume and macroscopic design but a more hydrophobic nature captured only ≈33%. An optimized adenovirus retention of 98% is achieved by adjusting the microchannels of the tunable inserts.

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An efficient capture strategy for the purification of human adenovirus type 5 from cell lysates

Cited by 6 publications

References 34 publications

Bridging upstream and downstream for improved adenovirus 5 bioprocess

Bridging upstream and downstream for improved adenovirus 5 bioprocess

Experimental study on the optimum number of layers of fiber mats for gas-liquid coalescer

3D‐Printed Highly Porous Functional Materials for the Efficient Removal of Adenovirus

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