Overcoming Downstream Purification Challenges for Viral Vector Manufacturing: Enabling Advancement of Gene Therapies in the Clinic

  title={Overcoming Downstream Purification Challenges for Viral Vector Manufacturing: Enabling Advancement of Gene Therapies in the Clinic},
  author={Orjana Terova and Steven M Soltys and Pim W Hermans and Jessica D. de Rooij and Frank J. M. Detmers},
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… Expand
Comparison of Different Liquid Chromatography-Based Purification Strategies for Adeno-Associated Virus Vectors
This work explored fast, scalable, and universal liquid chromatography-based strategies for the purification of rAAVs and found that anion exchange chromatography worked well for the AAV8 serotype and achieved high levels of purification and a baseline separation of full and empty rAAV. Expand
Moving from the bench towards a large scale, industrial platform process for adeno‐associated viral vector purification
Current scalable purification techniques that can be employed during the commercial manufacturing of AAV are summarized to highlight certain development considerations, such as adventitious agent removal and process development using the principals of quality by design. Expand
Upstream and downstream solutions for AAV manufacturing
With the advancement of gene delivery vectors and gene editing technologies, cell and gene therapies are a very real solution to many previously untreatable or difficult to treat diseases. With thisExpand
Peptides and pseudopeptide ligands: a powerful toolbox for the affinity purification of current and next-generation biotherapeutics.
This work presents a comprehensive and systematic review of the literature on peptide-based ligands and their use in the affinity purification of established and upcoming biological drugs. Expand
A high cell density perfusion process for Modified Vaccinia virus Ankara production: Process integration with inline DNA digestion and cost analysis
A detailed cost analysis comparing integrated production in batch versus production in perfusion mode showed that the cost per dose for MVA was reduced by nearly one‐third using this intensified small‐scale process. Expand
The COVID-19 Vaccine Race: Challenges and Opportunities in Vaccine Formulation
This review described the COVID-19 vaccines that are currently tested in clinical trials and provided in-depth insight into the various types of vaccines, their compositions, advantages, and potential limitations. Expand
Therapeutic Fc fusion protein misfolding: A three-phasic cultivation experimental design
The results showed that by utilizing two-step temperature shift, productivity and the quality of target protein have been improved simultaneously and this model could be successfully applied to other products. Expand
Affinity Chromatography: An Enabling Technology for Large‐Scale Bioprocessing
This mini‐review is to provide an overview of the history of bioprocess AC, the current state of platform processes based on affinity capture steps, the maturing field of custom developed biobrocess affinity resins, and the advantages of affinity capture‐based downstream processing in comparison to other forms of chromatography. Expand
Super-resolution optical microscopy reveals direct virus binding at hybrid core-shell matrices.
In the present study, for the first time the viral binding behavior at virus-imprinted polymers (VIPs) via stimulated emission depletion (STED) microscopy is shown by imaging individual, fluorescently labeled virus particles. Expand
The Impact of Product and Process Related Critical Quality Attributes on Immunogenicity and Adverse Immunological Effects of Biotherapeutics.
A quality target product profile (QTPP) that identifies the key CQAs through process development can help assign severity scores based on safety, immunogenicity, pharmacokinetics (PK) and pharmacodynamics (PD) of the molecule. Expand


Scalable Downstream Strategies for Purification of Recombinant Adeno-Associated Virus Vectors in Light of the Properties
This review will evaluate the scalable downstream purification strategies systematically, especially those based on the physicochemical properties of AAV virus, and attempt to find better scalable downstream strategies for rAAV vectors. Expand
Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale.
A high-yielding, recombinant AAV production process based on polyethylenimine-mediated transfection of HEK293 cells and iodixanol gradient centrifugation of concentrated culture supernatant is described, demonstrating equivalent or better transduction both in vitro and in vivo when compared with small-scale, CsCl gradient-purified vectors. Expand
Manufacturing of recombinant adeno-associated viral vectors for clinical trials
This chapter will provide a description of the current methods to produce clinical grade of rAAV under current good manufacturing practice (cGMP) settings. Expand
AAV Biology, Infectivity and Therapeutic Use from Bench to Clinic
This chapter reviews current information on rAAV clinical trials and the potential for combining rAAv platform with other technologies, such as induced pluripotent cells and gene editing. Expand
AAV hybrid serotypes: improved vectors for gene delivery.
This review summarizes the studies performed on AAV retargeting, and categorizes the available hybrid serotypes to date, based on the type of modification: 1) transcapsidation, 2) adsorption of bi-specific antibody to capsid surface, 3) mosaic capsid, and 4) chimeric capsid. Expand
Production, purification and characterization of adeno-associated vectors.
The current state of the art in the various production and purification methods for rAAVs is reviewed, including classical parameters and methodologies to characterize rAAV stocks will be discussed. Expand
A simplified baculovirus-AAV expression vector system coupled with one-step affinity purification yields high-titer rAAV stocks from insect cells.
A method is described for the production of rAAV serotypes 1 and 2 in insect cells using a simplified baculovirus-AAV expression vector system coupled with particle purification via affinity chromatography. Expand
Enabling Platform Approaches to Advance Gene Therapies
O ver the past decade, gene therapy applications and their importance in the biopharmaceutical industry have been increasing. Gene therapies promise versatile treatment options that couldExpand
Viral vectors for gene therapy: Current state and clinical perspectives
This review summarizes the latest trends in theDevelopment of gene therapy, in particular, various aspects and prospects of the development of clinical products based on viral delivery systems. Expand
Undetectable transcription of cap in a clinical AAV vector: implications for preformed capsid in immune responses.
R residual cap expression was undetectable following transduction with AAV2-hFIX clinical vectors, and performed capsid protein is implicated as the source of epitopes recognized by CD8(+) T cells that eliminated vector-transduced cells in the clinical study. Expand