Moving from the bench towards a large scale, industrial platform process for adeno‐associated viral vector purification

2020 ◽  
Vol 117 (10) ◽  
pp. 3199-3211
Author(s):  
Benjamin Adams ◽  
Hanne Bak ◽  
Andrew D. Tustian
Keyword(s):  
Large Scale ◽  
Viral Vector ◽  
Platform Process

scholarly journals Antigen Presentation of mRNA-Based and Virus-Vectored SARS-CoV-2 Vaccines

Vaccines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 848
Author(s):  
Ger T. Rijkers ◽  
Nynke Weterings ◽  
Andres Obregon-Henao ◽  
Michaëla Lepolder ◽  
Taru S. Dutt ◽  
...  
Keyword(s):  
Antigen Presentation ◽  
Conformational Changes ◽  
Dna Sequences ◽  
Neutralizing Antibodies ◽  
Large Scale ◽  
Viral Vector ◽  
Adenovirus Vector ◽  
Platelet Factor ◽  
Immune Mediated ◽  
Protein Encoding

Infection with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) causes Coronavirus Disease 2019 (COVID-19), which has reached pandemic proportions. A number of effective vaccines have been produced, including mRNA vaccines and viral vector vaccines, which are now being implemented on a large scale in order to control the pandemic. The mRNA vaccines are composed of viral Spike S1 protein encoding mRNA incorporated in a lipid nanoparticle and stabilized by polyethylene glycol (PEG). The mRNA vaccines are novel in many respects, including cellular uptake and the intracellular routing, processing, and secretion of the viral protein. Viral vector vaccines have incorporated DNA sequences, encoding the SARS-CoV-2 Spike protein into (attenuated) adenoviruses. The antigen presentation routes in MHC class I and class II, in relation to the induction of virus-neutralizing antibodies and cytotoxic T-lymphocytes, will be reviewed. In rare cases, mRNA vaccines induce unwanted immune mediated side effects. The mRNA-based vaccines may lead to an anaphylactic reaction. This reaction may be triggered by PEG. The intracellular routing of PEG and potential presentation in the context of CD1 will be discussed. Adenovirus vector-based vaccines have been associated with thrombocytopenic thrombosis events. The anti-platelet factor 4 antibodies found in these patients could be generated due to conformational changes of relevant epitopes presented to the immune system.

scholarly journals 277. Large Scale rVSV Viral Vector Production in a cGMP Facility

2010 ◽  
Vol 18 ◽  
pp. S105-S106
Keyword(s):  
Large Scale ◽  
Viral Vector

scholarly journals Multivalent and Multipathogen Viral Vector Vaccines

2016 ◽  
Vol 24 (1) ◽  
Author(s):  
Katharina B. Lauer ◽  
Ray Borrow ◽  
Thomas J. Blanchard
Keyword(s):  
Immune Response ◽  
Measles Virus ◽  
Viral Vectors ◽  
Large Scale ◽  
Viral Vector ◽  
Natural Infection ◽  
Scale Production ◽  
Veterinary Vaccine ◽  
Large Scale Production ◽  
Foreign Proteins

ABSTRACT The presentation and delivery of antigens are crucial for inducing immunity and, desirably, lifelong protection. Recombinant viral vectors—proven safe and successful in veterinary vaccine applications—are ideal shuttles to deliver foreign proteins to induce an immune response with protective antibody levels by mimicking natural infection. Some examples of viral vectors are adenoviruses, measles virus, or poxviruses. The required attributes to qualify as a vaccine vector are as follows: stable insertion of coding sequences into the genome, induction of a protective immune response, a proven safety record, and the potential for large-scale production. The need to develop new vaccines for infectious diseases, increase vaccine accessibility, reduce health costs, and simplify overloaded immunization schedules has driven the idea to combine antigens from the same or various pathogens. To protect effectively, some vaccines require multiple antigens of one pathogen or different pathogen serotypes/serogroups in combination (multivalent or polyvalent vaccines). Future multivalent vaccine candidates are likely to be required for complex diseases like malaria and HIV. Other novel strategies propose an antigen combination of different pathogens to protect against several diseases at once (multidisease or multipathogen vaccines).

Preparation of a recombinant adeno-associated viral vector with a mutation of human factor IX in large scale and its expressionin vitro andin vivo

2001 ◽  
Vol 46 (16) ◽  
pp. 1367-1371
Author(s):  
Huazhong Lu ◽  
Li Chen ◽  
Xuefeng Wang ◽  
Daru Lu ◽  
Xinfang Qiu ◽  
...  
Keyword(s):  
Large Scale ◽  
Human Factor ◽  
Viral Vector ◽  
Factor Ix ◽  
Human Factor Ix

scholarly journals Integrase-Defective Lentiviral Vectors for Delivery of Monoclonal Antibodies against Influenza

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1460
Author(s):  
Zuleika Michelini ◽  
Judith M. Minkoff ◽  
Jianjun Yang ◽  
Donatella Negri ◽  
Andrea Cara ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Influenza A ◽  
Large Scale ◽  
Viral Vector ◽  
Direct Injection ◽  
Lentiviral Vectors ◽  
Attractive Alternative ◽  
Scale Production ◽  
Safety Features

Delivering rapid protection against infectious agents to non-immune populations is a formidable public health challenge. Although passive immunotherapy is a fast and effective method of protection, large-scale production and administration of monoclonal antibodies (mAbs) is expensive and unpractical. Viral vector-mediated delivery of mAbs offers an attractive alternative to their direct injection. Integrase-defective lentiviral vectors (IDLV) are advantageous for this purpose due to the absence of pre-existing anti-vector immunity and the safety features of non-integration and non-replication. We engineered IDLV to produce the humanized mAb VN04-2 (IDLV-VN04-2), which is broadly neutralizing against H5 influenza A virus (IAV), and tested the vectors’ ability to produce antibodies and protect from IAV in vivo. We found that IDLV-transduced cells produced functional VN04-2 mAbs in a time- and dose-dependent fashion. These mAbs specifically bind the hemagglutinin (HA), but not the nucleoprotein (NP) of IAV. VN04-2 mAbs were detected in the serum of mice at different times after intranasal (i.n.) or intramuscular (i.m.) administration of IDLV-VN04-2. Administration of IDLV-VN04-2 by the i.n. route provided rapid protection against lethal IAV challenge, although the protection did not persist at later time points. Our data suggest that administration of mAb-expressing IDLV may represent an effective strategy for rapid protection against infectious diseases.

scholarly journals Development of the VIGS System in the Dioecious Plant Silene latifolia

2019 ◽  
Vol 20 (5) ◽  
pp. 1031 ◽  
Author(s):  
Naoko Fujita ◽  
Yusuke Kazama ◽  
Noriko Yamagishi ◽  
Kyoko Watanabe ◽  
Saki Ando ◽  
...  
Keyword(s):  
Sex Determination ◽  
Candidate Genes ◽  
Large Scale ◽  
Viral Vector ◽  
Functional Characterization ◽  
Virus Vector ◽  
Functional Gene ◽  
Silene Latifolia ◽  
Vector System ◽  
Dioecious Plant

(1) Background: Silene latifolia is a dioecious plant, whose sex is determined by XY-type sex chromosomes. Microbotryum lychnidis-dioicae is a smut fungus that infects S. latifolia plants and causes masculinization in female flowers, as if Microbotryum were acting as a sex-determining gene. Recent large-scale sequencing efforts have promised to provide candidate genes that are involved in the sex determination machinery in plants. These candidate genes are to be analyzed for functional characterization. A virus vector can be a tool for functional gene analyses; (2) Methods: To develop a viral vector system in S. latifolia plants, we selected Apple latent spherical virus (ALSV) as an appropriate virus vector that has a wide host range; (3) Results: Following the optimization of the ALSV inoculation method, S. latifolia plants were infected with ALSV at high rates in the upper leaves. In situ hybridization analysis revealed that ALSV can migrate into the flower meristems in S. latifolia plants. Successful VIGS (virus-induced gene silencing) in S. latifolia plants was demonstrated with knockdown of the phytoene desaturase gene. Finally, the developed method was applied to floral organ genes to evaluate its usability in flowers; (4) Conclusion: The developed system enables functional gene analyses in S. latifolia plants, which can unveil gene functions and networks of S. latifolia plants, such as the mechanisms of sex determination and fungal-induced masculinization.

scholarly journals Thrombotic events and COVID-19 vaccines

2021 ◽  
Vol 25 (9) ◽  
pp. 701-707
Author(s):  
C. Brazete ◽  
A. Aguiar ◽  
I. Furtado ◽  
R. Duarte
Keyword(s):  
Immune Thrombocytopenia ◽  
Large Scale ◽  
Viral Vector ◽  
Vaccine Safety ◽  
Severe Thrombocytopenia ◽  
The European Union ◽  
Vein Thrombosis ◽  
Post Marketing Surveillance ◽  
Post Marketing ◽  
Vaccination Campaigns

COVID-19 vaccines are considered promising agents in the control of the pandemic. Although their safety was assessed in randomised clinical trials, severe adverse events (AEs) have been reported after large-scale administration. This study aims to evaluate thromboembolic AEs reported after vaccination in a real-world context and how they led to the interruption of vaccination campaigns. We also review the benefits and risks of the vaccines approved in the European Union and provide recommendations. A review of the literature was performed using Medline/PubMed electronic database as well as institutional and pharmacovigilance official reports. Our findings show that vaccine-induced prothrombotic immune thrombocytopenia has been suggested as a very rare AE associated with viral vector vaccines. Unusual thrombotic events combined with moderate-to-severe thrombocytopenia were reported mainly in women under 60 years of age. As safety signals emerged, Vaxzevria and Janssen´s COVID-19 vaccine campaigns have been paused while investigations proceed. On the other hand, the number of deep vein thrombosis and pulmonary embolism reports have not increased. Post-marketing surveillance indicated that mRNA vaccines are safe and should continue to be used. The thrombotic events report rate is not increased in people over 60 years. As they are at greater risk for COVID-19 complications and death, no vaccine restrictions are recommended in this group. Risk factors for vaccine-induced prothrombotic immune thrombocytopenia should be established so that evidence-based decisions can be made. Systematic monitoring of COVID-19 vaccine safety is essential to ensure that the benefits of vaccination outweigh the risks.

scholarly journals Adeno-associated and lentiviral vector production in 2D and 3D formats with adherent cells in chemically defined, blood-free media

2021 ◽  
Author(s):  
Sofia Pezoa ◽  
Randall Alfano ◽  
Atherly Pennybaker ◽  
Nathan Hazi ◽  
Andrew Laskowski
Keyword(s):  
Bovine Serum ◽  
Viral Vectors ◽  
Large Scale ◽  
Viral Vector ◽  
Fetal Bovine Serum ◽  
Adherent Cells ◽  
Scale Production ◽  
Fetal Bovine ◽  
Free Media ◽  
Doubling Times

Large scale manufacturing of viral vectors or vaccines with adherent cells still relies heavily on the inclusion of fetal bovine serum for the growth and production phases. The inclusion of serum presents numerous problems with the undefined chemical makeup, the undesirable safety profile, and the constraints and limitations on the global supply. Despite these challenges, alternatives to serum for adherent cells have been limited; however, advances in large-scale production of recombinant human proteins have enabled the advancement of blood-free media that can support adherent cell growth. In order to circumvent the need for serum in adherent platforms, we developed a serum and blood-free, chemically defined medium specific for adherent human epithelial kidney cells and evaluated growth kinetics as well as viral vector production with associated adenovirus and lentivirus. We observed doubling times equal to or faster than doubling times observed in serum containing medium. We also demonstrate transfection efficiencies and viral titers that are equivalent to or higher than that of serum. Our results demonstrate that fetal bovine serum is not required for culture of adherent HEK cells, and that a serum-free, blood-free, chemically defined approach can be reliably implemented in the production of viral vectors for gene therapy.

scholarly journals Monobac System–A Single Baculovirus for the Production of rAAV

2021 ◽  
Vol 9 (9) ◽  
pp. 1799
Author(s):  
Lionel Galibert ◽  
Aurélien Jacob ◽  
Adrien Savy ◽  
Yohann Dickx ◽  
Delphine Bonnin ◽  
...  
Keyword(s):  
Large Scale ◽  
Viral Vector ◽  
Cell System ◽  
Lower Amount ◽  
Efficient Production ◽  
Baculovirus Genome ◽  
Raav Vector ◽  
System A ◽  
Raav Production

Large-scale manufacturing of rAAV is a bottleneck for the development of genetic disease treatments. The baculovirus/Sf9 cell system underpins the first rAAV treatment approved by EMA and remains one of the most advanced platforms for rAAV manufacturing. Despite early successes, rAAV is still a complex biomaterial to produce. Efficient production of the recombinant viral vector requires that AAV replicase and capsid genes be co-located with the recombinant AAV genome. Here, we present the Monobac system, a singular, modified baculovirus genome that contains all of these functions. To assess the relative yields between the dual baculovirus and Monobac systems, we prepared each system with a transgene encoding γSGC and evaluated vectors’ potency in vivo. Our results show that rAAV production using the Monobac system not only yields higher titers of rAAV vector but also a lower amount of DNA contamination from baculovirus.

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