Biomedical Engineering and Bioengineering Commons^™

Development Of Recombinant Protein Based Chemical Conjugate Malaria Vaccines Targeting The Pre-Erythrocytic Stage, Transmission Blocking, Or Both, David L. Narum, Nicholas Macdonald, David Jones, Ruth Ellis, Yimin Wu, Patrick E. Duffy

Vaccine Technology III

The development of a Plasmodium falciparum malaria vaccine is critical for future control and elimination programs. Recombinant protein based chemical conjugate vaccines, covering different parasite stages, are being developed due to complexity of the parasite and sub-optimal immunogenicity of recombinant malaria proteins in humans, respectively. Chemical conjugation of recombinant malaria proteins to carrier proteins improves their immunogenicity in animal studies. A transmission blocking vaccine comprised of the ookinete protein Pfs25 chemically conjugated to Pseudomonas aeruginosa ExoProtein A (EPA) is currently being developed for pilot scale cGMP production. Bulk lots of Pfs25 and EPA have already been produced and released following …

Manufacturing Flu Vaccine In Mexico: A Major Public Health And Technology Transfer Challenge, Roger Vinas

Vaccine Technology III

As Health partner of Mexican Public authorities initiative, Sanofi Pasteur is committed to build a Flu Bulk Vaccine Site, on Mexican soil, in order, for this country, to be self-sufficient for supplying seasonal vaccine and, especially,in case of a Flu Pandemic event. This presentation will give highlights on key drivers to consider while transferring this kind of Technology from Europe to Mexico: - Technology Transfer guidelines: stepwise process - Getting Permits/Authorizations - Looking for partners/Contractors: Engineering and Construction phases - Staffing ramp-up for Manufacturing (Production and Quality Control) start-up

Design Of Experiment Based Japanese Encephalitis Virus Formaldehyde Inactivation Optimisation For A Vero Cell Derived Vaccine, Michael Hughson

Vaccine Technology III

Japanese Encephalitis (JE) is a disease primarily dominant in South East Asia, caused by the Japanese Encephalitis virus (JEV). It is responsible for an estimated 50,000 cases of the disease every year, of which around 10,000 are fatal and approximately 15,000 result in long term neurological sequelae (WHO 2006). Viral inactivation is a main feature in many vaccine manufacturing processes in order not only to inactivate the product itself but also any potential adventitious agents which may have been introduced during manufacture. Chemical inactivation using formaldehyde is one of the most common methods used for inactivated viral vaccines, yet due …

New Vaccine Technologies: Promising Advances May Save More Lives, John Boslego

Vaccine Technology III

In the past 20 years, immunization has prevented nearly 20 million deaths from vaccine-preventable infections. Despite this success, poorer countries often lack access to newer and more expensive vaccines, and vaccines are not yet available for many illnesses. PATH is working to narrow the immunization gap between developed countries and developing countries by increasing the availability of existing vaccines, reducing the lag time for adoption of recently-licensed vaccines, developing technology in support of vaccines and immunization (e.g. vaccine vial monitor, Uniject, vaccine stabilization platforms) and working with partners to develop new vaccines.

Vaccine development is expensive and manufacturers often focus …

Ultra-Scale Down Studies Of Human Cell Bioprocessing For A Prostate Cancer Vaccine Therapy - The Impact Of Capillary Shear, Juan Pablo Acosta Martinez, Katherine Lawrence, Carol Chu Mike Hoare, Stephen Ward

Vaccine Technology III

The scale-up and manufacturing of therapies based on intact whole cells presents a major challenge for development scientists and engineers due to the stress-reactive nature of these cells. The administrated cells may be characterized in terms of their membrane integrity and their surface markers and eventually their biopotency. The challenge is to process the cells at various scales and in a way which maintains these cell properties. Also during formulation the presence of cytokines produced by cells prior to their inactivation is a critical factor. This poster presents an approach to allow the rapid characterization of human cell lines in …

Escherichia Coli Plasmid Dna Fermentation: Strain And Process-Specific Effects On Vector Yield, Quality, And Transgene Expression, Aaron E. Carnes, Jeremy Luke, Justin Vincent, Clague Hodgson, James Williams

Vaccine Technology III

To commercialize DNA medicines, industrial plasmid DNA manufacturing processes are needed which meet the quality, economy, and scale requirements projected for future products. NTC has developed an inducible fed-batch fermentation process that incorporates novel cell bank and fermentation process innovations that reduce plasmid mediated metabolic burden. This process also incorporates a scalable plasmid induction profile that, in combination with vector backbone modifications that double fermentation productivity compared to existing high copy vectors such as pVAX1 and gWIZ, form a generic plasmid DNA production platform driving high plasmid yields up to 2.6 g/L, with specific yields of 5% of total dry …

Development Of Protein Capsular Matrix Vaccine (Pcmv) Technology, Kevin P. Killeen, Ann Thanawastien, Tom Griffin

Vaccine Technology III

Matrivax R&D Corp. is a start-up biotechnology company with R&D operations located in Boston, USA and a vaccine pilot facility in Haikou, China. We are developing a proprietary vaccine process that entraps polysaccharides in a cross-linked protein ‘carrier’ or matrix, termed Protein Capsular Matrix Vaccine (PCMV), as an alternative to conjugate vaccine technology. Despite highly efficacious pneumococcal vaccines such as Prevnar®, S. pneumoniae causes > 1 million deaths worldwide annually. Likewise, typhoid fever afflicts ~16 million people, resulting in 600,000 deaths despite effective vaccines such as Typhim Vi® and Ty21a. The premise is that inexpensive,efficacious polysaccharide vaccines that elicit TH-cell ‘memory’ …

Process Scale-Up And Optimization For Production Of High Efficacy Oral Rabies Vaccine, Amine Kamen, Chun Fang Shen, Stephane Lanthier, Danielle Jacob, Johnny Montes, Andrew Beresford

Vaccine Technology III

Rabies is an important causative agent of disease resulting in an acute infection of the nervous system and death of the individual. Rabies remains an important public health program in developing countries, and the indigenous threat of rabies continues in developed countries because of wildlife reservoirs. Globally, there are about 55,000 fatal human cases of rabies each year [WHO, 2007]. Control of rabies in wildlife remains an important challenge for government offices.

There are numbers of rabies vaccines commercially available for controls of wildlife rabies. However, these vaccines currently distributed to wildlife do not effectively immunize all at-risk species, especially …

Production Of Serotype 6-Derived Recombinant Adeno-Associated Virus In Serum-Free Suspension Cultures Of Hek 293 Cells, Érica A. Schulze

Vaccine Technology III

Recombinant adeno-associated virus (rAAV) has become a promising candidate vector for gene therapy. Anchorage-dependent cells are traditionally used to produce rAAV. However, mass production to meet demands for clinical trials requires large-scale and cost-effective manufacturing processes. The key advantages of rAAV vectors are a broader tissue tropism through the use of different serotypes and a good safety profile.

In the present work, a serotype 6-derived rAAV was produced by transfection of suspension HEK293 cells in serum-free medium using three plasmids: one encoding the GFP gene flanked by ITR sequences, the second encoding the replication and capsid genes, and a third …

Rna Based Plasmid Selection System For Antibiotic-Free Plasmid Dna Vector Production, Aaron E. Carnes, Jeremy Luke, Justin Vincent, Clague Hodgson, James Williams

Vaccine Technology III

Antibiotic resistance markers, typically kanamycin resistance (kanR), allow selective retention of plasmid DNA during bacterial fermentation and are the most commonly utilized selectable markers. However, to ensure safety, regulatory agencies recommend elimination of antibiotic resistance markers from therapeutic and vaccine plasmid DNA vectors. The presence of an antibiotic resistance gene in the plasmid backbone is considered undesirable by regulatory agencies, due to: 1) the potential transfer of antibiotic resistance to endogenous microbial flora; and 2) the potential activation and transcription of the genes from mammalian promoters after cellular incorporation into the genome. Here, we describe the development and application of …

Influence Of Host Cell Defence During Influenza Vaccine Production In Mdck Cells, Timo Frensing, Claudius Seitz, Bjoern Heynischotto-Von-Guericke, Udo Reichl

Vaccine Technology III

For cell culture-based influenza vaccine production virus yield optimisation is of crucial importance. In particular, with the recent threat of the new H1N1 pandemic, not only seasonal vaccines but also pre-/pandemic vaccines have to be supplied in large quantities. In vivo influenza replication is limited by the immune system, but for production cell lines the impact of cellular defence mechanisms on virus yield is unknown. In influenza-infected adherent Madin-Darby canine kidney (MDCK) cells the interferon (IFN) response and subsequent induction of the antiviral state was monitored. Virus yield and host cell signalling intensity were strain-dependent. By over-expression of viral antagonists …

Potent, Rapid And Cost-Effective Influenza Vaccines Made In E. Coli, Thomas Hofstaetter

Vaccine Technology III

The traditional influenza vaccine, trivalent inactivated virus (TIV) has been in use in the United States in one form or another since the 1940s. The system for production involves injecting influenza virus into embryonated hen’s eggs, harvesting the allantoic fluid containing the virus, inactivation with formalin, disruption of the virus with non-ionic detergent, zonal centrifugation to enrich for antigen and a second inactivation step. The recent appearance of the H1N1 swine virus has provided an opportunity to test the pandemic response system established over the past ten years. What we find is that the public health system seems to work …

Scale-Up Of An Intensified Process For Rad35 Adenovirus Production Using The Per.C6® Cell Substrate, Alfred Luitjens, Herman Van Herk

Vaccine Technology III

Tuberculosis is the world’s second deadliest infectious disease, with over 9 million new cases diagnosed in 2006. In collaboration with the Aeras Global Tuberculosis Vaccine Foundation, we are developing a recombinant tuberculosis adenovirus based vaccine (rAd35). Currently a series of three Phase I trials are taking place, with the first two studies indicating very promising results, safety and toleration. In November 2008 we started a Phase II study in South Africa. The manufacturing process supporting these clinical studies was developed using the PER.C6® cell substrate. With the productivity of this production process, scale-up to 10,000-liter bioreactor will be required to …

The Challenge Of Developing New Generation Vaccines For Control And Eradication Of Foot And Mouth Disease In South America, Susana Levy

Vaccine Technology III

Foot and mouth disease (FMD) is a highly infectious viral disease that affects food producing animals such as cattle, pigs and sheep. The FMD status given by the World Organization for Animal Health (OIE) has a huge financial impact on countries that have economies based on meat trade. In the past 15 years the MERCOSUR countries (Argentina, Brazil, Paraguay and Uruguay) have consolidated industrial processes to supply the region with safe and efficacious vaccines to prevent FMD in livestock. Control and eradication programs rely heavily on compulsory vaccination with multivalent whole virus inactivated vaccines. Currently, vaccines are industrially obtained by …

A Vaccine Prototype Using Baculovirus Expression System For The Control Of Avian Influenza Virus, Mauricio Realpe, R. Mora, L. Castellanos, F. Robles, C. González-Hernández

Vaccine Technology III

A clade 1 sequence of H5 haemaglutinin from an Asian Avian H5N1 isolate was used as a the template to chemically synthetize a codon optimized version for expression in insect cells. A single clone was chosen for expression optimization and changes were introduced in order to maximize the amount of protein to be produced and to resemble another sequence demonstrated to be present in an isolate causing disease in Humans.

Preliminary analysis at lab scale have shown promising yields of the haemaglutinin using an activity titration assay and an ELISA-based detection method. Optimization of the cell seeding, MOI, and time …

Vaccine Stabilization – Research, Commercialization, And Impact, Ray Cummings, Debra Kristensen, Dexiang Chen, Michel Zaffran

Vaccine Technology III

Improving the stability of vaccines and removing vaccines from refrigerated storage for all or part of their shelf life have been proposed as strategies to help ensure vaccine effectiveness and to mitigate cold chain storage capacity constraints and escalating costs associated with the introduction of new vaccines into developing countries. Over the last six years, PATH has conducted research on stabilization of measles, Haemophilus influenzae type b (Hib), hepatitis B, enterotoxgenic Escherichia coli, conjugate meningococcal A, and pentavalent (Diphtheria-Tetanus-Pertussis-hepatitis B-Hib) vaccines in collaboration with 22 technical partners and 10 vaccine development groups. Results will be shared and include marked improvement …

Purification Process Development Of Protein Subunit Based Vaccine Candidates Produced Using Recombinant E. Coli Expression System (Part - Ii), Davinder Chawla, Yan-Ping Yang

Vaccine Technology III

Proteins are key components of prophylactic vaccines against infectious diseases. Protein subunit based vaccine is an attractive alternative to the traditional detoxified bacterial or inactivated viral vaccine approach due to its highly purified and well characterized product nature. Purification of protein antigens to achieve consistent product purity and quality is an integral part of the protein subunit vaccine product development process. Expression levels of the recombinant proteins in bacterial expression system may be extremely high following rapid technology advancement. The challenges and approaches used to develop purification processes for novel protein vaccine candidates expressed at g/L level will be discussed.

Iscomatrix™ Adjuvant Links Innate And Adaptive Immune Responses, Debbie Drane

Vaccine Technology III

The ISCOMATRIX™ adjuvant has antigen delivery and presentation properties as well as immunomodulatory capabilities which combine to provide enhanced and accelerated immune responses. The responses are broad, including a range of sub classes of antibodies as well as both CD4+ and CD8+ T cells. A range of ISCOMATRIX™ vaccines (ISCOMATRIX™ adjuvant combined with antigen) have now been tested in clinical trials and have been shown to be generally safe and well tolerated as well as immunogenic, generating both antibody and T cell responses.

The mechanisms by which ISCOMATRIX™ adjuvant facilitates its immune effects is the scope of significant study and …

Development Of A Multi-Dose Formulation For Prevnar 13™, Lakshmi Khandke, Cindy Yang, Jingrun Fan, Hanyoung Han, Ksenia Krylova Abbas Rashidbaigi, Bruce A. Green, Kathrin U. Jansen

Vaccine Technology III

Streptococcus pneumoniae causes up to a million cases per year of invasive disease in young children and infants, most occurring in developing countries. Pfizer is partnering with WHO and the GAVI alliance to support the developing world on immunization against pneumococcal disease. Prevnar 13™ is currently approved for the prevention of invasive pneumococcal disease in twenty seven countries. If used widely, this pneumococcal conjugate vaccine could prevent hundreds of thousands of additional cases of child mortality each year.

Use of single-dose preservative-free vaccine formulations will raise the overall cost of vaccination programs and may jeopardize the effectiveness of immunization programs …

Purification Process Development Of Protein Subunit Based Vaccine Candidates Produced Using Recombinant E. Coli Expression System (Part - I), Davinder Chawla, Yan-Ping Yang

Vaccine Technology III

Proteins are key components of prophylactic vaccines against infectious diseases. Protein subunit based vaccine is an attractive alternative to the traditional detoxified bacterial or inactivated viral vaccine approach due to its highly purified and well characterized product nature. Purification of protein antigens to achieve consistent product purity and quality is an integral part of the protein subunit vaccine product development process. Expression levels of the recombinant proteins in bacterial expression system may be extremely high following rapid technology advancement. The challenges and approaches used to develop purification processes for novel protein vaccine candidates expressed at g/L level will be discussed.

Final Conference Program, Barry Buckland, John Aunins, Paula Marques Alves, Kathrin Jansen

Vaccine Technology III

List of talks during the conference.

Nipah/Hendra: Understanding The Links Between Human And Veterinary Emerging Diseases, Jules Minke

Vaccine Technology III

Animals constitute an important source of infectious diseases for humans and the majority of recent emerging diseases in humans are zoonotic. Infections occur through direct or indirect transmission from wildlife reservoirs or via the food chain. Nipah and Hendra viruses have recently joined the growing list of viruses that emerged from bats to cause serious disease in humans and livestock. Although the precise mode of virus transmission is not fully understood, human infection appears to be the result of close contact with infected horses and pigs that act as amplifying hosts. Improved control strategies are necessary to reduce the transmission …

After The License Approval: How Analytics Can Keep You In The Market, Robert Sitrin

Vaccine Technology III

Getting a BLA approved for a new vaccine can be a daunting task, but keeping the market supplied after licensure of a successful vaccine can be an even bigger challenge. Analytics can play an important role in keeping product supplied to the marketplace, especially during the critical launch phase The talk will describe the application of analytical comparability to GARDASIL®, Merck’s novel new vaccine to prevent cervical cancer, in order to bridge product produced by a launch facility to that produced by a scale-up facility without the need for a clinical trial. This effort assured a smooth transition of supply …

Establishing A Platform For Spray Drying Inhalable Vaccines In South Africa, Willem Germishuizen, L Venter, A Khosa, F Mudau, P.B. Fourie, B Pulliam, M Kabadi, A Schiermeier, D.A . Edwards

Vaccine Technology III

Mycobacterium bovis BCG is the current vaccine for tuberculosis (TB). However, BCG as it is currently administered shows highly variable efficacy in protecting adults against TB. The natural route of infection of TB is via inhalation of bacilli-containing aerosols and it is postulated that immunization by the natural route of infection may lead to a greater immunity given the fact that the lungs are the primary target of infection. By eliciting both local and systemic immune responses, it is anticipated that an inhaled form of BCG will offer greater protection against pulmonary TB.

Current commercial BCG vaccine preparations are filled …

Innovative Therapeutic Cancer Vaccines In Cuba: An Update, Luis Enrique Fernández

Vaccine Technology III

Worldwide cancer affects more than 10 million new people each year while other 22 million are living with this devastating disease.

Cancer vaccines are the newest weapon in the battle against this disease, specifically increasing body's natural abilities to halt tumour progression. Currently, 141 distinct projects are at different stages of clinical development.

The Cuban Biotech System of Institutions has been actively involved in this field for almost 20 years, resulting that 8 innovative therapeutic cancer vaccine projects are currently under clinical investigation.

The main focus of these projects is to impact the principal cancer localizations afecting Cuban population, like …

Application Of Animal-Free Recombinant Bioactive Protein Supplements To Improve The Performance Of Cell-Based Viral Vaccine Production, Kenneth Bertram, Marina Ross, Domenica Cavallaro, Larissa Chirkova, Geoffrey Francis

Vaccine Technology III

Animal cell culture for the production of viral vaccines has been performed for more than 50 years, and currently this technology is expanding rapidly to meet present and future demands of the health sector. The development and regulatory approval of continuous cell lines for manufacturing viral vaccines has brought numerous benefits to production processes. However, greater advances in the last decade have been achieved in mammalian cell production of biological therapeutics, including monoclonal antibodies, hormones, growth factors, cytokines and clotting factors. We and others have contributed to these upstream advances by improving cell culture media with the development of animal-free …

Building Process Understanding For Vaccine Manufacturing Using Data Mining, Matthew Wiener

Vaccine Technology III

The production of vaccines is a complex biological process, with long cycle times and a high level of variation in raw materials, biological growth rates, and test methods. While long-term shifts or cycles in yield are not unusual, it is important to build understanding of the causes of shifts and cycles, for greater control and predictability. Hundreds of variables are monitored for every batch of vaccine produced; however, the relationships between product quality and the many process variables are difficult to quantify. In this article, we describe how mining historical process data using random forests and partial least squares (PLS) …

Rabies Virus Glycoprotein (Rvgp) Expression In Drosophila S2 Cells And By Semliki Forest Virus (Sfv). Synthesis And Protection Studies, Carlos Pereira

Vaccine Technology III

Here we report the utilization of Drosophila melanogaster Schneider 2 (S2) cells and the Semliki Forest Virus (SFV) expression system to produce the rabies virus glycoprotein (RVGP). Although they represent different expression systems, they offer similar advantages in generating high-level expression of functional membrane proteins. They are both relatively easy and safe to handle and are scalable. RVGP synthesized by S2 cells and SFV carrying an mRNA coding for RVGP (SFV-RVGP) were assayed in mouse protection studies. We have constructed S2 gene vectors with the hygromycin selection gene (H) in which the RVGP gene is inserted under the control of …

Cim Technology: Enabeling Economic Vaccine Purification, Matjaz Peterka, Franci Smrekar, Tony Brazzale, Marko Banjac, Petra Kramberger, Milos Barut, Ales Podgornik, Ales Strancar

Vaccine Technology III

Vaccines are diverse and complex biological molecules, complexes and particles. Different production technologies are used for vaccines manufacturing and every production process require somekind of vaccine purification. Purification of vaccines is technically challenging and was traditionally inefficient and partially neglected due to different economical and technical reasons. Density gradient ultracentrifugation, introduced in1960s is still a major purification step for many vaccines present on the market. As a complementary techniques, cross flow filtration and chromatography has been used. Conventional chromatography supports designed for protein purification have relatively small pore sizes with restricted access for large molecules and viruses. In addition mass …

Development Of Inactivated Polio Vaccine From Attenuated Sabin Strains For Clinical Studies And Technology-Transfer Purposes, Yvonne E. Thomassen, Wilfried A.M. Bakker, Janny Westdijk, Aart G. Van ‘T Oever, Nico Van Den Heuvel, Jan Hendriks, Gideon F.A. Kersten, Leo A. Van Der Pol

Vaccine Technology III

Recently, responding to WHO’s call for new polio vaccines, the development of Sabin-IPV (injectable, formalin-Inactivated Polio Vaccine, based on attenuated ‘Sabin’ polio virus strains) was initated at NVI. This activity plays an important role in the WHO polio eradication strategy. The use of Sabin instead of wild-type Salk polio strains will provide additional safety during vaccine production. Initially, the Sabin-IPV production process will be based on the scale-down model of the current, and well-established, Salk-IPV process. In parallel, process development, optimization and formulation research is being carried out to further modernize the process and reduce cost per dose. The lab-scale …