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Expert Insight

Regulatory pathway on the manufacture & quality control of recombinant adeno-associated virus vectors

Spotlight Article

Expert Insight

Terence R Flotte & Allison M Keeler-Klunk

Latest Developments in Viral & Non-Viral Vector Manufacturing

Human gene therapy vectors are rapidly emerging as a new class of licensed products for single gene disorders. Examples include a retroviral gene therapy of hematopoietic stem cells for severe combined immune deficiency due to adenosine deaminase deficiency (ADA-SCID), marketed as Strimvelis® in Europe, and a recombinant adeno-associated virus (rAAV) vector for inherited retinal dystrophy due to RPE65 deficiency (IRD), approved as Luxturna® in the USA. The regulatory regime surrounding the human use of investigational human gene therapy products in the USA and Europe has been developed in the context of multiple trials of these vectors in patients over the past 28 years. Aspects of this framework relevant for rAAV vectors are discussed here.

Submitted for peer review: Jan 19 2018 Published: Mar 14 2018
DOI: 10.18609/cgti.2018.012
Citation: Cell Gene Therapy Insights 2018; 4(2), 89-99.
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Overcoming Downstream Purification Challenges for Viral Vector Manufacturing: Enabling Advancement of Gene Therapies in the Clinic

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Innovator Insight

Orjana Terova, Stephen Soltys, Pim Hermans, Jessica De Rooij & Frank Detmers

Latest Developments in Viral & Non-Viral Vector Manufacturing

The adeno-associated vector (AAV) has shown great potential for the delivery of therapeutic genes in the gene therapy field and has become a vector of choice for many therapies. Although excellent clinical outcomes have been reported, the current potential of viral vectors in the clinic is limited due to manufacturing challenges including the absence of an efficient and scalable platform purification process. Affinity chromatography has proven to be a viable solution for the purification of viral vectors and to date, a small number of, mostly single serotype-targeted affinity resins exist. However, to make these treatments economically viable, the ability to use one resin to purify multiple serotypes is vital. The POROSTM CaptureSelectTM AAVX chromatography resin can purify multiple AAV subclasses, including recombinant and chimeric vectors, with high binding efficiency. By reducing purification steps and maximizing productivity, this true pan-tropic AAV affinity resin offers a cost-effective purification solution for commercial AAV manufacturing.

Submitted for peer review: Dec 13 2017 Published: Mar 14 2018
DOI: 10.18609/cgti.2018.017
Citation: Cell Gene Therapy Insights 2018; 4(2), 101-111.
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Downstream bioprocessing of AAV vectors: industrial challenges & regulatory requirements

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Expert Insight

Matthias Hebben

Latest Developments in Viral & Non-Viral Vector Manufacturing

With an increasing number of clinical trials and impressive results in the field of gene therapy, recombinant adeno-associated virus (rAAV) is confirmed as one of the most promising viral vectors. Five years after the European Medicines Agency approved Glybera, the recent marketing authorization of Luxturna by the Food and Drug Administation in the United States marks the concretization of years of vector development. It also highlights the urgency to set up industrial manufacturing processes to support economically feasible commercialization. While several expression systems coexist as upstream processes in rAAV manufacturing, the downstream part is more conserved between processes. However, as there are no established specifications for rAAV vector preparation purity, it is not clear how to determine whether a purification process meets the safety requirements. In addition, there are characteristics unique to rAAV, like the presence of empty viral particles, which represent a technical hurdle to develop standardized and scalable downstream process leading to consistent quality of the vector. This review describes the existing purification technologies and the way they address current regulatory requirements.

Submitted for peer review: Jan 25 2018 Published: Mar 14 2018
DOI: 10.18609/cgti.2018.016
Citation: Cell Gene Therapy Insights 2018; 4(2), 131-146.
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Addressing scaling-up limitations: optimized PEI-mediated production of clinical grade viral vectors

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Innovator Insight

Alengo Nyamay’Antu, Valérie Kedinger & Patrick Erbacher

Latest Developments in Viral & Non-Viral Vector Manufacturing

With the progress in developing new viral vector systems guided by safety, specificity and potency considerations, several gene and cell-based therapies are now close to being clinically approved and commercially available to treat genetic diseases. These viral vectors systems are based on the production of mainly adeno-associated viruses (AAV) and lentiviruses by transient transfection of human embryonic kidney (HEK)-293 derived producer cell lines. Virus vector production using the right transient transfection method is crucial to provide the flexibility and reproducibility that is needed to scale-up from initial process development to manufacturing of high-quality grade viral vectors. PEI-mediated transient transfection is an efficient and cost-effective approach to produce viral vectors. With the available quality grades of PEI, PEIpro® and PEIpro-HQ®, Polyplus-transfection offers a reliable PEI transfection method suitable for process development up to viral vector production for clinical and commercial manufacturing.

Submitted for peer review: Dec 15 2017 Published: Feb 22 2018
DOI: 10.18609/cgti.2018.013
Citation: Cell Gene Therapy Insights 2018; 4(2), 71-79.
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Addressing the Challenges of Commercial-Scale Viral Vector Production

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Innovator insight

Dr Nicole Faust

Latest Developments in Viral Vector Manufacturing

Dr Nicole Faust has extensive management experience in the biotechnology sector. Over the last 18 years she has held scientific management positions with Artemis/Taconic Biosciences, amaxa, Lonza, and CEVEC, engaging in the fields of pharmaceutical models, gene transfer, protein production, and gene therapy. She received her Ph.D. from University of Freiburg and an MBA degree from Educatis University, Switzerland.

DOI: 10.18609/cgti.2018.004
Citation: Cell Gene Therapy Insights 2018; 4(2), 31-36.
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Analytical approaches to characterize AAV vector production & purification: Advances and challenges

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Expert Insight

Fabien Dorange & Christine Le Bec

Latest Developments in Viral & Non-Viral Vector Manufacturing

Adeno-associated viral (AAV) vectors have been reported to be a great promise in a large number of clinical trials. Multiple AAV gene products will enter into early and late phase clinical trials. To sustain this, reliable, fast, robust, GMP compliant analytical methods and characterization protocols are needed. Specific analytical assays need to be performed to assess vector productivity, vector purity, biological activity and safety. Besides the potency of the vector, current advances appear to improve the methods for the quantification and characterization of AAV particles that are empty or contain DNA fragments other than the full vector genome of interest. This article describes the analytical methods available to measure the strength/dose of the AAV vector, to quantify full/empty particles and illegitimate encapsidated DNA, their limitations and recent improvements.

Submitted for peer review: Jan 29 2018 Published: Mar 14 2018
DOI: 10.18609/cgti.2018.015
Citation: Cell Gene Therapy Insights 2018; 4(2), 119-129.
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Merits of non-viral cellular engineering versus viral cellular engineering

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Innovator insight

Dr Jessica Carmen

Latest Developments in Viral & Non-Viral Vector Manufacturing

Dr Carmen is the Director of Strategic Marketing for Cellular Therapy at MaxCyte, Inc. MaxCyte is driving the next generation of cell-based medicines with its best in-class cell modification technology which is used in the discovery, development, and manufacture of small molecule, biologic, and cell-based medicines. She is responsible for the growth of partnerships with key opinion leaders and developers of ex vivo-modified cellular therapies. Dr Carmen joined MaxCyte in 2016. In her previous roles she was responsible for establishing partnerships with developers of cell-based medicines in order to expand a cell therapy contract manufacturing business. Dr Carmen has been instrumental in the establishment of the Standards Coordinating Body for Regenerative Medicines and is a member of the Board of Directors. She is an active member of the Science and Technology Committee of the Alliance for Regenerative Medicine and has served on the Commercialization Committee for the International Society of Cellular Therapy. Dr Carmen earned a Doctorate from the Johns Hopkins University School of Public Health and a Bachelor of Science from the University of Florida.

DOI: 10.18609/cgti.2018.018
Citation: Cell Gene Therapy Insights 2018; 4(2), 113-117.
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