Digital Commons Network^™

Production Of Monoclonal Antibodies Specific To Soluble Proteins Of Bartonella Bacilliformis, Yanina Zarate Sulca, Yolanda Medina, Karen D. Calvay Sanchez, Giovanna Mendonza Mujica

Vaccine Technology VIII

Bartonella bacilliformis is the causative agent of Carrion’s disease, this illness affects poor communities of Andean valleys (Peru, Colombia and Ecuador) [1]. B. bacilliformis produces a biphasic infection: acute phase presents severe anemia and immunosuppression, and chronic phase involves dermal eruption [1]. Currently, the available tools for the diagnosis of Carrion’s disease include low sensitive serological assays and expensive molecular tools [2]. To improve the diagnosis, this project aims to produce monoclonal antibodies specific soluble proteins of B. bacilliformis. The monoclonal antibodies production was performed immunizing three Balb/C mice with soluble proteins obtained from the total lysate of B. …

Design Of A Vaccine Against Dengue And Zika Viruses Based On A Mimotope Of The Envelope Dimer Epitope, Esmeralda Cuevas-Juárez, Arturo Liñan, Carolina Hernandez, Mykhailo Kopylov, Octavio T. Ramírez, Laura A. Palomares

Vaccine Technology VIII

Zika and dengue viruses are members of the Flavivirus genus that cause mild fever, rash and general body pain; but can cause severe reactions, as hemorrhages (dengue virus), congenital syndrome (Zika virus), or even death. Because of the structural similarity between these viruses, some antibodies generated after an infection can cross-react with different members of the flavivirus family. After a secondary infection, the cross-reactive antibodies can lead to more severe forms of the disease, through a mechanism named antibody-dependent enhancement of infection (ADE). Broadly neutralizing antibodies are antibodies that neutralize both, dengue and Zika viruses; and it has been demonstrated …

Generation Of An International Standard Serum To Measure Influenza Virus Hemagglutinin Stalk-Reactive Antibodies, Juan Manuel Carreno Quiroz, Tara Hurst, Raffael Nachbagauer, Mohammad Amin Behzadi, Shirin Strohmeier, Lynda Coughlan, Othmar Engelhardt, Florian Krammer

Vaccine Technology VIII

Current efforts towards the development of a universal influenza virus vaccine rely on induction of effective long-term antibody responses against conserved regions of the influenza virus glycoproteins. The stalk domain of the hemagglutinin has been targeted for this purpose due to its high degree of conservation among numerous influenza subtypes and strains. Several vaccine candidates targeting this domain are in late pre-clinical or early clinical stage of development. Currently assays to measure stalk-based immunity are not standardized and no international standard is available. As various vaccine developers are generating data from pre-clinical and clinical studies, harmonization of assay read-outs will …

Intensified High Yield Production Of Gmma Based Vaccines For High Burden Neglected Disease, Luigi Sollai, Angela Daniele, Giulia Iannello, Federico Pippi, Antonia De Felice, Francesca Necchi, Rocio Cannals, Francesco Berlanda Scorza, Anna Maria Colucci

Vaccine Technology VIII

Vaccines are among the most important public health interventions currently available. The global demand for vaccines is growing due to global population growth, ongoing global immunization campaigns and the increase of antibiotic-resistant bacteria. Vaccines are of high importance in Low- and Middle-Income Countries (LMIC) where millions of deaths occur annually due to vaccines-preventable diseases. Global coverage would need at least 200 million doses annually of a two-dose vaccine to cover the at-risk birth cohort, and considerably more for catch up vaccination during a roll out phase. Especially for vaccines against diseases in LMIC, there is a major added constraint: the …

Lessons In Vaccine Process Development, Tara Tagmyer

Vaccine Technology VIII

The biopharmaceutical industry mounted an impressive response to the rapidly evolving COVID-19 pandemic by reprioritizing pipelines and establishing novel collaborations. While most biopharmaceutical companies attempted to develop a vaccine or treatment, it was the companies that previously invested in more platform-based technologies, such as mRNA and viral vectors, that tended to respond more quickly to the evolving situation. As a result, multiple safe and efficacious vaccines were authorized for emergency use, resulting in significant declines in severe disease and mortality. However, durable immunity to COVID-19 has not yet been demonstrated. While the rapid development and deployment of efficacious vaccines is …

Influenza A Virus-Derived Defective Interfering Particles For Antiviral Treatment, Sascha Y. Kupke, M D. Hein, Y Genzel, L Pelz, P Marichal-Gallardo, N Bdeir, S Pöhlmann, U Rand, L Cicin-Sain, F Alnaji, C Brooke, M Winkler, L Cicin-Sain, D Bruder, K Schughart, U Reichl

Vaccine Technology VIII

Here, we report on genetically engineered, propagation-incompetent influenza A virus (IAV) particles, so-called defective interfering particles (DIPs) that have been suggested as a promising novel antiviral agent. Typically, IAV DIPs harbor a large internal deletion in one of their eight genomic viral RNA (vRNA) segments. Further, DIPs are capable of hijacking cellular and viral resources upon co-infection with fully infectious standard virus (STV), resulting in an antiviral effect. Besides this replication interference, DIP infection also stimulates innate immunity, adding to the antiviral efficacy.

So far, DIPs were produced in embryonated chicken eggs. To improve scalability and flexibility of processes as …

Production Of High-Quality Sars-Cov-2 Antigens For Vaccine Development And Serological Assays Implementation, Bárbara Fernandes, Rute Castro, Patricia Gomes-Alves, Mario Amacker, Toon Stegmann, Sylvain Fleury, Paula M. Alves, António Roldão

Vaccine Technology VIII

The development of novel and/or improved vaccines as well as the establishment of tools to monitor vaccine responses are two key factors to control COVID-19 pandemic, often requiring the manufacturing of significant amounts of high-quality SARS-CoV-2 antigens.

In this work, we produced SARS-CoV-2 Spike (S) and the receptor binding domain (RBD) proteins in human or insect cell lines to be further used in (i) the implementation of serological assays for detection of antibodies against SARS-CoV-2 virus in the Portuguese population, and (ii) the development of a virosome-based COVID-19 vaccine candidate.

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Advances In Bioprocessing, Analytics And Formulation Of Influenza Ha-Vlp Vaccine Candidates Produced By Insect Cells, António Roldão, Ricardo Correia, Bárbara Fernandes, Sofia B. Carvalho, Ricardo J. S. Silva, Cristina Peixoto

Vaccine Technology VIII

The emergence of new influenza strains demands the continued development of novel, flexible, and scalable platforms for vaccine production. In this study, we describe advancements in the manufacturing process of influenza hemagglutinin (HA)-displaying virus-like particle (VLP)-based vaccines produced by insect cells, from upstream and downstream processing to analytics and formulation.

Aiming to improve influenza HA-VLPs production, evolutionary engineering and process intensification have been applied. Adaptation of stable Sf-9 cells producing HA-VLPs to hypothermic growth resulted in up to 12-fold higher expression. Likewise, adaptation of parental High Five cells to neutral pH induced a 3-fold higher specific HA-VLPs production rate following …

Conference Program, Francesc Godia, Linda Hwee-Lin Lua, Charles Lutsch, Tarit Mukhopadhyay

Vaccine Technology VIII

No abstract provided.

Development Of The Oxford Astazeneca Covid Vaccine, Sarah Gilbert

Vaccine Technology VIII

The 2014 outbreak of Ebola virus disease in West Africa, chiefly in Guinea, Sierra Leone, and Liberia, but with cases spreading to other countries highlighted the lack of preparedness for combating infectious disease outbreaks. The virus was first identified in 1976, but vaccine development had proceeded slowly and no candidate vaccines had progressed further than phase I trials. In 2014 two new candidate vaccines entered rapid clinical development with encouraging early results, but efficacy trials were slow to start. Ebola is only one of many known viruses with the potential to cause outbreaks, and with the support of the WHO …

On-Line Influenza Virus Quantification For Viral Production Processes Thanks To Affinity-Based Surface Plasmon Resonance Biosensor, Emma Petiot, Laurent Durous, Manuel Rosa-Calatrava, Christophe A. Marquette

Vaccine Technology VIII

Influenza virus seasonal epidemics, associated with the constant threat of new pandemic outbreak, challenge vaccine manufacturers to develop responsive processes that can outreach the limitations of traditional egg-based technology. Recent progress made regarding cell culture bioprocesses allowed for numerous alternative strategies to developed future vaccine candidates, as for example the recombinant HA or Virus—like Particles (VLP) vaccines. However, while cell culture allows for more versatility than ovoculture, regarding process development and monitoring, these alternatives still require optimization to seriously concurrence the traditional process. To drive these developments, WHO and regulatory agencies underlined the need for developing better influenza vaccine potency …

A Rapid Response Vaccine Manufacturing Platform As A Countermeasure To Epidemic Threats, Tania Pereira Chilima, Héla Kallel

Vaccine Technology VIII

The preparedness and rapid response to emerging infectious diseases outbreaks remains challenging due to limited market incentives. The development and manufacturing of vaccines are today facing bottlenecks in development time as well as in cost-effectiveness in order to be able to react rapidly and deliver low-cost, high-quality vaccines.

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The Next Generation Of Fibroblast-Based Vaccine Development, Anna-Barbara Hachmann, Megan Pajak-Lee, David Klinkenberg, Andrew M. Campbell, Aneg L. Cortes, Isabel M. Gimeno

Vaccine Technology VIII

Chicken embryo fibroblasts (CEF) and diploid cells have a long history in vaccine production since their isolation in the 1960s at the Wistar Institute (WI-38 cells) as well as the Medical Research Council (MRC-5 cells). The cells quickly became adopted for a number of vaccines: varicella zoster (VZV), MMR, yellow fever, polio, hepatitis A, rotavirus, rabies, Marek's disease, and dengue virus. Most of these vaccine processes were developed with classical media supplemented with Fetal Bovine Serum (FBS). The Hayflick limit of diploid cells restricted their adaptation to a serum-free process. While some of the vaccines such as polio and rabies …

The Implication Of Glycans On The Ace2: Sars-Cov-2 Spike Interaction, Manuel Reithofer, Rong Zhu, Yoo Jin Oh, Reingard Grabherr, Peter Hinterdorfer, Miriam Klausberger

Vaccine Technology VIII

Since its emergence in 2019 the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) continues to a profoundly impact and threaten human health. For the development of novel prophylactic and therapeutic measures a detailed understanding of the virus-host interaction and features that modulate the interaction is of utmost importance. Attachment of the SARS-CoV-2 virus to human host cells predominantly relies on the specific interaction of the viral spike (S) surface glycoprotein with the receptor angiotensin-converting enzyme 2 (ACE-2). Glycans within or surrounding the binding interface have been demonstrated to play an important role in the ACE2:S interaction. The quality of this …

Differential Glycosylation And Extracellular Vesicle Biogenesis In Hek293 Upon Transient Transfection, Jesús Lavado-García, Irene González-Domínguez, Laura Cervera, Inmaculada Jorge, Jesús Vázquez, Tao Zhang, Manfred Wuhrer, Francesc Gòdia

Vaccine Technology VIII

Vaccine therapies based on virus-like particles (VLPs) are currently increasing relevance due to the strong immune response they elicit and their manufacture advantages when compared to traditional biopharmaceuticals. During VLP production using mammalian cell-based platforms, different extracellular vesicles (EVs) are coproduced, leading to the need of a complex downstream purification process. Currently there is no effective and efficient method to separate VLPs from EVs which share very similar density and physicochemical properties^1,2. Different methods to characterize the EV composition and their protein content and glycosylation signature are used in this work to further understand their biochemical nature. First, …

Biomanufacturing Using Single Use Systems: Case Study Of Fluoropolymer Material, Julien Muzard, Muhammad Siddiqui, Ross Acucena

Vaccine Technology VIII

In this presentation, we will discuss recent biocompatibility data obtained using our gamma stable fluoropolymer platform and its advantage in a modern manufacturing environment for the handling of almost any biological fluids.

Single-use, disposable solutions are now widely accepted as gold standard in the biopharmaceutical industry covering every single steps of the drug manufacture process. From early-stage small scale R&D project, upstream production with several thousand-liters bioreactors, downstream process, API formulation up to critical fill & finish and transport applications, the industry is taking a shift with a significant breakthrough reported in 2021: the largest GMP facility in the world …

Use Of A Parallel Bioreactor Scaledown System For Optimisation Of A Perfusion-Based Upstream Process For Adenovirus Production, Carina C.D. Joe, Steffi Scholze, Julia Niemann, Alexander D. Douglas

Vaccine Technology VIII

The recent pandemic emphasises the need for vaccine producers to be able to respond rapidly to the need for large quantities for global distribution. Here, we report work to optimise a perfusion-based upstream approach. Perfusion can enhance volumetric productivity of adenoviral vectors, but the complexity of perfusion culture and the lack of suitable scale-down models has hindered work to establish the complex relationships between variables affecting the process.

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Driving Change In Dtap Batch Release Testing, Isabelle Bekeredjian-Ding, Bärbel Friedrichs, Walter Matheis, Björn Becker, Christel Kamp, Olga Ticha, Volker Öppling

Vaccine Technology VIII

The complexity of vaccine manufacturing has raised the need to drive standardization and quality control requirements as well as batch release of vaccines. The purpose of release testing is to ensure that efficacy and safety of the vaccine product are maintained in all batches. Classical testing includes challenge experiments in animals that provide proof of vaccine potency and identify subpotent vaccines. However, novel concepts such as “consistency testing” question the continued need for in vivo experiments and propose to implement rigorous QC for lot-to-lot consistency testing with other methods at an earlier stage.

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Workshop: Process Intensification Considerations For Lmic, Philippe Alexandre Gilbert

Vaccine Technology VIII

Please click Additional File below for the presentation.

New Insights In Formaldehyde-Induced Detoxification Of The Tetanus Toxin: Chemical Modification Stoichiometry And Characterization Of Intra- And Inter-Molecular Cross-Links, Thierry Eynard, Sébastien Peronin, Amaud Salvador, Fabien Barbirato, Vincent Colombie, Céline Rocca, Nour Al Turihi, Catherine Jourdat, Jérôme Lemoine

Vaccine Technology VIII

The active ingredient of the tetanus vaccine, known as tetanus toxoid (TTD, 150 kDa), is prepared by inactivation of the tetanus toxin with formaldehyde. This chemical treatment determines the efficiency and innocuity of the vaccine. The detoxification chemistry has been qualitatively studied regarding the nature and positioning of some formaldehyde-induced modifications. However, neither the stoichiometry of the chemical modifications or intra- and inter-chain cross-linking have been yet characterized, despite being of great interest for the definitive elucidation of the detoxification mechanism as well as for future manufacturing follow-up. The aim of the present study is to assess the impact of …

Virus Like Particles Expressed In Insect Cells And Mammalian Cells As A Plataform For The Development Of A Zika Vaccine, Selene J. Uribe, Ana Ruth Pastor, Diego Fontana, Claudio Prieto, Octavio T. Ramírez, Laura A. Palomares

Vaccine Technology VIII

The Zika virus (ZIKV) is an emergent mosquito-borne virus of the flaviviridae family that has caused severe challenges to global health since 2015. It causes Guillain-Barré syndrome and congenital malformations. Although case numbers have decreased, it is important to develop a vaccine for outbreaks. One alternative is the use of virus like particles (VLP) as vaccines. ZIKV is enveloped and is composed of three main structural proteins: enveloped (E), pre-membrane (M), and capsid. The main target of neutralizing antibodies is the E glycoprotein, which is glycosylated in some strains. The N-glycosylation profile is determined by the producer host cell. ZIKV …

Optimization, Production, Purification And Characterization Of Hiv-1 Gag Vlps Functionalized With Sars-Cov-2 Spike Glycoprotein, Amau Boix-Besora, Elianet Lorenzo, Jesus Lavado-Garcia, Francesc Gòdia, Laura Cervera

Vaccine Technology VIII

Virus-like particles (VLPs) constitute a promising approach for recombinant vaccine development. They are robust, safe, versatile and high immunogenic supra-molecular structures that closely mimic the native conformation of the viruses without carrying its genetic material. HIV-1 Gag VLPs share similar characteristics with wild type Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) virus, making them a suitable platform to express the Spike membrane protein to generate a potential vaccine candidate for COVID-19. This work proposes a methodology for the generation of SARS-CoV-2 VLPs by its co-expression with HIV-1 Gag protein by transient transfection of HEK 293 cultures. We first evaluated the …

Development Of A Pentavalent Group B Streptococcus (Gbs) Glycoconjugate Vaccine In Africa, Seanette Wilson, Ebrahim Mohamed, Patrick Tippoo, Matthew Williams

Vaccine Technology VIII

A vaccine against Group B Streptococcus (GBS), utilized in a maternal immunization strategy to reduce the burden of GBS mortality and morbidity in infants, has potential for use all over the world, but would have the greatest impact in low-resource countries that have limited access to interventions. Currently there is no vaccine against GBS. The Biovac Institute (Biovac), a private public partnership in South Africa, is developing a pentavalent, polysaccharide-protein conjugate vaccine (PCV) against GBS — using state-of-the-art technology and targeting the most common strains.

A vaccine providing more than 90 % coverage against GBS infection would need to include …

Multivalent Influenza Vaccine Production In Hek-293 Cells In Response To Pandemic Threats, Amine Kamen, Cécile Toussaint, Omar Farnos, David Sharon, Alina Venereo-Sanchez, Irene González-Domínguez, Alice Bernier, Xingge Xu, Hanan Chaabane

Vaccine Technology VIII

Influenza virus infects millions of people every year worldwide, with elderly and very young people among the most critically affected. Strains that constitute a pandemic threat are characterized by the severity of the clinical manifestations and mortality rates and tend to require the urgent production of hundreds of millions of vaccine doses in very short periods of time. There is an evident need to develop new generations of influenza vaccines based on robust production systems such as mammalian or insect cell cultures. These systems may allow, in contrast to production in embryonated chicken eggs, a faster response capacity, a superior …

Affinity Purification Of Sars-Cov-2 Spike Protein Receptor Binding Domain Produced In A C1 Fungal Expression System, Amit Dutta, Aditya Utturkar, Ronen Tchelet, Noelia Valbuena Crespo, Laura Mueller, Renaud Jacquemart

Vaccine Technology VIII

The Receptor Binding Domain (RBD)of the spike protein of SARS-CoV-2 has shown promise for diagnosis, treatment, and development of vaccines for COVID-19. However, two problems persist with large scale production of RBD: 1) lack of high productivity upstream cell culture, 2) absence of a commercial, highly selective affinity resin. In an effort to overcome these limitations, we evaluated two novel technologies for the production and purification of RBD.

Briefly, RBD was expressed using C1, an engineered fungal strain of Thermothelomyces heterothallica (DyadicInternational¹). The C1 platform expresses glycosylated antigens with high productivity, stability, and purity. RBD was purified using …

Accelerating And Intensifying Manufacturing To Enable Large-Scale Supply Of A New Adenovirus-Vectored Vaccine Within 100 Days, Sandy Douglas, Alexander D Douglas, Carina C D Joe, Rameswara R Segireddy, Adam Berg, Yuanyuan Li, Dimitrio Doultsinos, Cathy Oliveira, Asma Ahmad, Nitin Chopra, Steffi Scholze, Piergiuseppe Nestola, Julia Niemann

Vaccine Technology VIII

The Coalition for Epidemic Preparedness Innovations’ ‘100-day mission’ aspires to launch of a new vaccine within 100 days of pathogen identification. We have previously reported a simple fed batch process and strategy of internationally-distributed manufacturing, which enabled 2 billion doses of the ‘Oxford / AstraZeneca’ adenovirus-vectored COVID-19 vaccine to be produced in less than 600 days from publication of the SARS-CoV-2 genome sequence. The majority was made and used in low and middle income countries. Here, after briefly reviewing that previous work, we will describe efforts to further improve adenovirus manufacturing for response to future pathogen outbreaks and variants.

Please …

Multimodal Chromatography Combining Steric Exclusion And Cation Exchange As An Intermediate Downstream Step To Purify Yellow Fever Virus-Like Particles, Túlio M. Lima, Ricardo A. Medronho, Leda R. Castilho

Vaccine Technology VIII

Yellow fever (YF) is an hemorrhagic viral disease transmitted by infected mosquitoes, which is endemic in many African and Central/South American countries. The severe symptoms and the high mortality rate of the disease can have devastating effects in case an outbreak occurs in an area where the population is non-vaccinated. Before the current YF vaccine became available, outbreaks in cities like Barcelona (Spain) and Philadelphia (USA) led to the death of approximately 10% of the population. Recent outbreaks have shown that YF continues to be a major public health threat due to production capability issues and shortage of vaccine stockpiles, …

Influence Of Dna-Protein Interactions On Purification And Assembly Of Virus-Like Particles, Lukas Gerstweiler,, Jingxiu Bi, Linda Lua, Anton Middelberg

Vaccine Technology VIII

Modular virus-like-particles (VLPs), presenting foreign antigens on their surface, are promising candidates for a wide range of future vaccines. A viable production pathway is the expression and purification of viral structural proteins and their subsequent in vitro assembly into VLPs, in a bioprocess environment. One promising approach is the use of murine polyomavirus major capsid protein VP1 as a carrier of modular epitopes from vaccine targets. This platform technology uses E. coli as an expression system and showed promising results in creating VLP vaccines candidates directed at influenza, Group A Streptococcus and other infectious pathogens. However, like other viral capsomeres …

Increasing Of Pk15 Biomass For Pcv Viral Antigen Production Using Modelbased And Media Optimization Strategies That Consider Cellular Metabolic Requirements, Ziomara Gerdtzen, Felipe Véliz, Gerardo Flores, Felipe Sánchez, Andrea Villanueva, Kurt Pohlhammer, Daniel Morales, J. Cristian Salgado, Iván Valdés, Harold Oliva, Samuel Valdevenito

Vaccine Technology VIII

Porcine circovirus associated disease (PCVD) is responsible for a significant economic loss in the livestock industry. A vaccine based on a viral antigen has been developed to prevent this disease, and it is produced industrially in the pig’s kidney cell line PK15. However, both biomass and viral antigen production are limited in the current attachment culture bioprocess set-up.

Please click Download on the upper right corner to see the full abstract.

Tackling A Capacity Bottleneck To Permit Large-Scale Downstream Processing Of An Adenovirus-Vectored Vaccine, Carina Citra Dewi Joe, Thomas Linke, Piergiuseppe Nestola, Yuanyuan Li, Rameswara R. Segireddy, Richard Turner, Alexander D. Douglas

Vaccine Technology VIII

We recently described the strategy by which the University of Oxford and AstraZeneca collaboratively scaled up production of our adenovirus-vectored COVID-19 vaccine, using a productive fed batch process and distributed manufacturing approach in twelve countries around the world.

Here we will focus on the development of the downstream process used to make this vaccine. In early development, the first tangential flow filtration step in our previously developed process was noted to be a potential obstacle for scale-up beyond 200L. By removing this first tangential flow filtration step, we established a simple purification process capable of handling the increasing quantities and …