Viral vector production in the integrity® iCELLis® single-use fixed-bed bioreactor, from bench-scale to industrial scale

Lennaertz, Alexandre; Knowles, Shane; Drugmand, Jean-Christophe; Castillo, José

doi:10.1186/1753-6561-7-s6-p59

Cited by 20 publications

(10 citation statements)

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“…20 Cell growth can be measured using a permittivity-based biomass probe. 21 Diverse biologicals, including monoclonal antibodies, 22 vaccines, 19 , 23 gammaretroviral vectors, 24 adenoviral vectors, 25 adeno-associated viral vectors, 26 , 27 , 28 and LVVs 29 , 30 have been produced in the iCELLis reactor, but often with little or no optimization. With this wide array of applications, much need exists to investigate the effects of culture parameters in the iCELLis reactor.…”

Section: Introductionmentioning

confidence: 99%

Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor

Powers

Drury

Hoehamer³

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Self-inactivating lentiviral vectors (LVVs) are used regularly for genetic modification of cells, including T cells and hematopoietic stem cells for cellular gene therapy. As vector demand grows, scalable and controllable methods are needed for production. LVVs are typically produced in HEK293T cells in suspension bioreactors using serum-free media or adherent cultures with serum. The iCELLis® is a packed-bed bioreactor for adherent or entrained cells with surface areas from 0.53 to 500 m 2 . Media are pumped through the fixed bed and overflows, creating a thin film that is replenished with oxygen and depleted of CO 2 as media return to the reservoir. We describe the optimization and scale-up of the production of GPRTG-EF1α-hγ c -OPT LVV using a stable packaging cell line in the iCELLis Nano 2-cm to the 10-cm bed height low compaction bioreactors (0.53 and 2.6 m 2 surface area) and compare to the productivity and efficacy of GPRTG-EF1α-hγ c -OPT LVV manufactured under current Good Manufacturing Practice (cGMP) using 10-layer cell factories for the treatment of X-linked severe combined immunodeficiency. By optimizing fetal bovine serum (FBS) concentration, pH post-induction, and day of induction, we attain viral yields of more than 2 × 10 7 transducing units/mL. We compared transduction efficiency between LVVs produced from the iCELLis Nano and cell factories on healthy, purified CD34 + cells and found similar results.

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Section: Introductionmentioning

confidence: 99%

Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor

Powers

Drury

Hoehamer³

et al. 2020

Molecular Therapy - Methods & Clinical Development

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“…Uneven cell distribution in the iCELLis ® Nano (4 m 2 , 10 cm fixed-bed) might have some impact on total productivity. Data shows that the bioreactor supports the large scale adenovirus infection and production, and thus far iCELLis ® Nano has already shown to be suitable for several other products such as AAV (30), retrovirus (31), Rabies,…”

Section: Discussionmentioning

confidence: 98%

“…A novel fixed-bed bioreactor, the iCELLis ® provides a recent development providing a polyethylene terephthalate (PET) matrix for adherent cell growth. The iCELLis ® Nano has been used for a range of vector applications, such as for AAV (13), retrovirus (14), Rabies, Hepatitis-A and Chikungunya vaccine production (15). In this study, we evaluated for the first time the fixed bed iCELLis ® bioreactor for the manufacturing of Ad5 vectors in a HEK293 cell line.…”

Section: Introductionmentioning

confidence: 99%

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Lesch

Heikkilä

Lipponen³

et al. 2015

Human Gene Therapy

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Large-scale vector manufacturing for phase III and beyond has proven to be challenging. Upscaling the process with suspension cells is increasingly feasible, but many viral production applications are still applicable only in adherent settings. Scaling up the adherent system has proven to be troublesome. The iCELLis(®) disposable fixed-bed bioreactors offer a possible option for viral vector manufacturing in large quantities in an adherent environment. In this study, we have optimized adenovirus serotype 5 manufacturing using iCELLis Nano with a cultivation area up to 4 m(2). HEK293 cell cultivation, infection, and harvest of the virus (by lysing the cells inside the bioreactor) proved possible, reaching total yield of up to 1.6×10(14) viral particles (vp)/batch. The iCELLis 500 is designed to satisfy demand for large-scale requirements. Inoculating a large quantity of cell mass into the iCELLis 500 was achieved by first expanding the cell mass in suspension. Upscaling the process into an iCELLis 500/100 m(2) cultivation area cassette was practical and produced up to 6.1×10(15) vp. Flask productivity per cm(2) in iCELLis Nano and iCELLis 500 was in the same range. As a conclusion, we showed for the first time that iCELLis 500 equipment has provided an effective way to manufacture large batches of adenoviral vectors.

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“…[ 44 ] With fixed‐bed bioreactors, the lysis has been performed by traditional freeze‐and‐thaw cycles. [ 9,15 ] The process step is not very practical as one should freeze the whole bioreactor, or optionally unpack the bioreactor and remove the carriers in order to perform the freeze‐and‐thaw cycle for the carriers only. Unpacking creates an open system and is impractical to perform in a large scale.…”

Section: Virus Productionmentioning

confidence: 99%

Evaluation of the Single‐Use Fixed‐Bed Bioreactors in Scalable Virus Production

Lesch¹,

Valonen

Karhinen

2020

Biotechnology Journal

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The accelerating development of gene therapy from research towards clinical trials and beyond has elevated the demand for practical viral vector‐manufacturing solutions. The use of disposable upstream technology is gaining traction in clinical manufacturing. Packed‐bed or fixed‐bed reactors, where column is packed with immobilized biocatalyst particles providing surface to constrain the cells in a particular region of the reactor, have been widely used in bioprocessing applications since mid‐1900s. However, the world's first single‐use, fully integrated, high cell density, fixed‐bed bioreactor was launched only approximately a decade ago. By now, several single‐use, fixed‐bed technology solutions have been developed in a small scale. Scaling‐up the manufacturing can be challenging and for commercial‐scale manufacturing, there is practically only one single‐use, good manufacturing practice‐compliant option available. This study reviews the latest, fully disposable, fixed‐bed bioreactors; compares the virus production in the different systems; and discusses important manufacturing cost‐related topics. It is predicted that single‐use, fixed‐bed bioreactors will receive even more attention in the field of viral vector manufacturing and commercialization, especially with the need for higher virus titers and virus yields.

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Viral vector production in the integrity® iCELLis® single-use fixed-bed bioreactor, from bench-scale to industrial scale

Cited by 20 publications

References 0 publications

Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor

Lentiviral Vector Production from a Stable Packaging Cell Line Using a Packed Bed Bioreactor

Process Development of Adenoviral Vector Production in Fixed Bed Bioreactor: From Bench to Commercial Scale

Evaluation of the Single‐Use Fixed‐Bed Bioreactors in Scalable Virus Production

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