Lentiviral Vector Purification Using Nanofiber Ion-Exchange Chromatography

Ruščić, Jelena; Perry, Christopher; Mukhopadhyay, Tarit; Takeuchi, Yasu; Bracewell, Daniel G.

doi:10.1016/j.omtm.2019.08.007

Cited by 35 publications

(35 citation statements)

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“…For chromatography, the use of convective driven devices is effective in LV purification schemes and new stationary phases are emerging with very promising results. [ 36 ] Additionally, affinity chromatography using tailor‐made ligands might prove valuable due to its high selectivity allowing to decrease the number of purification steps and increasing recovery yields. [ 82,106 ]…”

Section: Discussionmentioning

confidence: 99%

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Advances in Lentivirus Purification

Moreira

Cavaco

Faria

et al. 2020

Biotechnology Journal

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Lentiviral vectors (LVs) have been increasingly used as a tool for gene and cell therapies since they can stably integrate the genome in dividing and nondividing cells. LV production and purification processes have evolved substantially over the last decades. However, the increasing demands for higher quantities with more restrictive purity requirements are stimulating the development of novel materials and strategies to supply the market with LV in a cost-effective manner. A detailed review of each downstream process unit operation is performed, limitations, strengths, and potential outcomes being covered. Currently, the majority of large-scale LV manufacturing processes are still based on adherent cell culture, although it is known that the industry is migrating fast to suspension cultures. Regarding the purification strategy, it consists of batch chromatography and membrane technology. Nevertheless, new solutions are being created to improve the current production schemes and expand its clinical use.

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Section: Discussionmentioning

confidence: 99%

“…The authors claimed to achieve a volumetric concentration factor of 100, with a recovery yield of 85%. [ 36 ]…”

Section: Scalable Methods For LV Downstream Processingmentioning

confidence: 99%

Advances in Lentivirus Purification

Moreira

Cavaco

Faria

et al. 2020

Biotechnology Journal

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“…Other development in the chromatographic field is the cellulose nanofibers, a new adsorbent design material that has an open structure large enough to be accessible to viral vectors, allowing mass transfer based on convection. Adenovirus type 5 and Lentiviral particles have been recently purified using this nanofiber-based ion-exchange chromatography [120,121]. The most recent purification processes for different biotherapeutic particles and their respective production system, purification steps and recovery yields are present in Table 1.…”

Section: Intermediate Purificationconcentration and Chromatographymentioning

confidence: 99%

Current challenges in biotherapeutic particles manufacturing

Moleirinho

Silva

Alves

et al. 2019

Expert Opinion on Biological Therapy

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Introduction: The development of novel complex biotherapeutics led to new challenges in biopharmaceutical industry. The potential of these particles has been demonstrated by the approval of several products, in the different fields of gene therapy, oncolytic therapy, and tumor vaccines. However, their manufacturing still presents challenges related to the high dosages and purity required. Areas covered: The main challenges that biopharmaceutical industry faces today and the most recent developments in the manufacturing of different biotherapeutic particles are reported here. Several unit operations and downstream trains to purify virus, virus-like particles and extracellular vesicles are described. Innovations on the different purification steps are also highlighted with an eye on the implementation of continuous and integrated processes. Expert opinion: Manufacturing platforms that consist of a low number of unit operations, with higheryielding processes and reduced costs will be highly appreciated by the industry. The pipeline of complex therapeutic particles is expanding and there is a clear need for advanced tools and manufacturing capacity. The use of single-use technologies, as well as continuous integrated operations, are gaining ground in the biopharmaceutical industry and should be supported by more accurate and faster analytical methods.

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“…Such phases are defined by thin electrospun threads by which the mobile phase can flow past and retention is on the fibres themselves. Such nanofibers have seen utility in the bind-and-elute of LV [216] and adenovirus [225], although commercial capacity is currently focused on the mAb market. Some hybrid mixed mode chromatography applications have also been developed.…”

Section: Chromatographic Purificationmentioning

confidence: 99%

“…Screening studies can provide crucial bioprocess information such as the level of transmembrane pressures [65] or crossflow rate [145] in TFF operations, column flow rate in AEX chromatography [145] or the right molecular weight cut-off (MWCO) or membrane material in TFF operation [65,145,243,244]. It is also important to determine the impact of using frozen-thawed materials in process development of unformulated LV products (e.g., [190,216]), as opposed to using fresh material, as this step may have a huge influence on the process performance rather than as result of the bioprocess operation itself. The fundamental study and understanding of LV bioprocessing and rational bioprocess development will be crucial to address this.…”

Section: Generationmentioning

confidence: 99%

Lentiviral Vector Bioprocessing

Perry

Rayat

2021

Viruses

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Lentiviral vectors (LVs) are potent tools for the delivery of genes of interest into mammalian cells and are now commonly utilised within the growing field of cell and gene therapy for the treatment of monogenic diseases and adoptive therapies such as chimeric antigen T-cell (CAR-T) therapy. This is a comprehensive review of the individual bioprocess operations employed in LV production. We highlight the role of envelope proteins in vector design as well as their impact on the bioprocessing of lentiviral vectors. An overview of the current state of these operations provides opportunities for bioprocess discovery and improvement with emphasis on the considerations for optimal and scalable processing of LV during development and clinical production. Upstream culture for LV generation is described with comparisons on the different transfection methods and various bioreactors for suspension and adherent producer cell cultivation. The purification of LV is examined, evaluating different sequences of downstream process operations for both small- and large-scale production requirements. For scalable operations, a key focus is the development in chromatographic purification in addition to an in-depth examination of the application of tangential flow filtration. A summary of vector quantification and characterisation assays is also presented. Finally, the assessment of the whole bioprocess for LV production is discussed to benefit from the broader understanding of potential interactions of the different process options. This review is aimed to assist in the achievement of high quality, high concentration lentiviral vectors from robust and scalable processes.

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Lentiviral Vector Purification Using Nanofiber Ion-Exchange Chromatography

Cited by 35 publications

References 35 publications

Advances in Lentivirus Purification

Advances in Lentivirus Purification

Current challenges in biotherapeutic particles manufacturing

Lentiviral Vector Bioprocessing

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