Download or read book Bioprocess Development for Plasmid based Vaccine Production written by Clarence Ongkudon and published by . This book was released on 2011 with total page 508 pages. Available in PDF, EPUB and Kindle. Book excerpt: Plasmid DNA (pDNA) vaccine is a promising vaccine technology, with better safety profile, more economical production and transport logistics, than conventional viral vaccines. Most importantly, pDNA vaccines elicit different immune responses including antibody-mediated, CD4 T-cell-mediated and CD8 T-cell-mediated immune responses, for defending against viral infections and cancer. The increasing number of preclinical and clinical trials on plasmid vaccines has triggered the need to make more in less time. Recent developments in theproduction of plasmid therapeutics involve the establishment of innovative and cost effective methods as well as simplified operations. This dissertation reports fundamental studies essential to the development of a rapid economically-viable plasmid production system which is cGMP-compatible. Optimisation of upstream bacterial fermentation and continuousdownstream purification of the plasmid vaccine fraction are the main aspects considered in the project of this dissertation. Process variables required to improve the volumetric and specific yields of a model plasmid-based measles vaccine (pcDNA3F) harboured in E. coli DH5[alpha] were investigated. A cGMP-compatible method offering the capacity to continuouslyproduce homogeneous supercoiled pDNA from clarified bacterial lysate using a monolithicadsorbent was developed. The method involved optimisation of the adsorbent characteristics, ligand functionalisation and chromatographic process conditions. The feasibility of using free metal ions to preferentially precipitate endotoxins (LPS) from a clarified plasmid DNAcontainingbacterial lysate was investigated. Screening of various free metal ions for effectiveendotoxin removal and optimisation of process conditions, such as pH, ion concentration, temperature and incubation time, using central composite design experiments were performed. The potential and advantages of using Zn2+-induced LPS aggregation as a secondary pDNA purification method was validated by studying the interaction of Zn2+ with LPS and pDNA. A comparative economic analysis on the basis of vaccine cost per dose for influenza vaccine produced via pDNA vaccine technology and fertilised egg-based technology was also studied. Experimental results from growth medium optimisation in 500 mL culture showed a maximum volumetric yield of 13.65 mg/L, twice the amount generated using a standard medium (PDM). Fed-batch fermentation in combination with exponential glycerol feeding strategy resulted in a significant increase of 110 mg/L pcDNA3F volumetricyield and a specific yield of 14 mg/g. In addition, growth pH variation (6 to 8.5) andtemperature fluctuation (35 oC to 45 oC) also resulted in improved plasmid yield.Chromatographic purification of pDNA using a triethylamine-activated conical monolithicadsorbent resulted in preferential pcDNA3F adsorption with optimum resolution achievedunder the conditions of 400 nm pore size of monolith, 0.7 M NaCl (pH 6) of binding bufferand 3 % B/min of gradient elution up to 1 M NaCl. Plasmid volumetric yield and recovery of ~3g/L and ~90% were obtained. Contaminant levels recorded were protein (0.01 mg/L), LPS(0.12 EU/mg) with no detectable gDNA and RNA. Results from endotoxin removal andanalysis showed that ZnSO4 displayed the highest endotoxin removal efficiency (~91%) and plasmid recovery (~100%). It was found that selective endotoxin precipitation ( 0.05 EU/Mg) could effectively be carried out during neutralisation in alkaline cell lysis at a pH condition similar to that of clarified cell lysate, a low ZnSO4 concentration (0.5 M), a minimum incubation time (30 min) and a temperature of 15 oC. Apparently, the lipopolysaccharide (LPS) showed a decreased aggregate size at the start of the ZnSO4 addition before increasing gradually. Results from the LPS aggregation analysis drew a hypothesis thatcationic close range encounter and interaction with LPS monomers may contribute to LPS self-aggregation whilst bridging of LPS monomers may increase the LPS aggregate size to a greater extent compared to that of self-aggregation. Specifically, addition of Zn2+ resulted in the largest number of LPS particles per aggregate and the value of aggregation constant (Km)for LPS-Zn2+ was substantially low (0.28 M) and considerably large (2 M) for pDNA-Zn2+,indicating its preferential ability to remove LPS from pDNA-containing solutions. The economic studies suggested that pDNA-based influenza vaccine production was highly dependent on the selling price and production volume. A similar cost per dose of about $2 was calculated although most of the manufacturing costs for plasmid DNA vaccine were lower than inactivated virus vaccine. This dissertation has developed a simple bioprocess framework to successfully improve production specification of plasmid vaccines using pcDNA3F as a model. The method offers ease of plasmid DNA purification due to reduced bulk impurities, cost-efficiency and most importantly high endotoxin removal (> 80%) and plasmid recovery (> 90% ). The technology will have a great impact on overall plasmidproduction and in particular on the development of axial flow monolithic purification in combination with selective endotoxin precipitation.
Download or read book Development of New Tools for the Production of Plasmid DNA Biopharmaceuticals written by Diana Morgan Bower and published by . This book was released on 2012 with total page 108 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA vaccines and gene therapies that use plasmid DNA (pDNA) as a vector have gained attention in recent years for their good safety profile, ease of manufacturing, and potential to treat a host of diseases. With this interest comes increased demand for high-yield manufacturing processes. Overall, this thesis aims to develop new, innovative tools for the production of plasmid DNA biopharmaceuticals. As one part of this thesis, we designed a 1-mL fed-batch microbioreactor with online monitoring and control of dissolved oxygen, pH, and temperature, as well as continuous monitoring of cell density. We used the microbioreactors to scale down temperature-induced production of a pUC-based DNA vaccine vector, pVAX1-GFP. Scaled-down processes can facilitate high-thoughtput, low-cost bioprocess development. We found that the microbioreactors accurately reproduced the behavior of a bench-scale bioreactor as long as key process parameters, such as dissolved oxygen, were held constant across scales. The monitoring capabilities of the microbioreactors also provided enhanced process insight and helped identify conditions that favored plasmid amplification. A second aspect of this thesis involved construction and characterization of a new DNA vaccine vector based on a runaway replication mutant of the R1 replicon. Runaway replication plasmids typically show increased amplification after a temperature upshift. However, we found that our new vector, pDMB02-GFP, gave higher yields during constant temperature culture at 30"C, reaching a maximum of 19 mg pDNA/g DCW in shake flasks. We gained mechanistic insight into this behavior by measuring RNA and protein expression levels of RepA, a plasmid-encoded protein required for initiation of replication at the R1 origin. Through these studies we found that RepA levels may limit plasmid amplification at 42*C, and relieved this limitation by increasing RepA translation efficiency via a start codon mutation. We also scaled up production of pDMB02-GFP at 300C from 50-mL shake flasks to 2-L bioreactors. Initial scale up efforts resulted in increased growth rate compared to the shake flasks, accompanied by very low plasmid yields. Decreasing the growth rate by limiting dissolved oxygen increased plasmid specific yield and emerged as a viable strategy for maintaining productivity during scale up.
Download or read book Bioprocessing of Viral Vaccines written by Amine Kamen and published by CRC Press. This book was released on 2022-09-06 with total page 267 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book focuses on cell culture-produced viral vaccines to meet the needs of the rapidly expanding research and development in academia and industry in the field. This book introduces the basic principles of vaccination and the manufacturing of viral vaccines. Bioprocessing of Viral Vaccines, will provide an overview of the advanced strategies needed to respond to the challenges of new and established viral infection diseases. The first few chapters cover the basics of virology and immunology as essential concepts to understand the function and design of viral vaccines. The core of the content is dedicated to process development, including upstream processing and cell culture of viral vaccines, downstream processing, and extensive analytical technologies specific to viral vaccines. Advanced process analytical technologies (PAT) and Quality by Design (QbD) concepts are also introduced in the context of vaccine manufacturing. The case studies included cover inactivated, attenuated vaccines exemplified by influenza vaccines, sub-unit vaccines exemplified by Virus Like Particles (VLPs: HPV vaccines) and sub-unit vaccines (Flublock), vectored vaccines: adenoviruses and Vesicular stomatitis Virus (VSV) vectored vaccines, genomic vaccines (DNA and mRNA) vaccines as developed for COVID-19 response in particular and a review of COVID-19 vaccines approved or in advanced clinical trials. This book is aimed at graduate engineers and professionals in the fields of vaccinology, bioprocessing, and biomanufacturing of viral vaccines.
Download or read book Vaccine Development and Manufacturing written by Emily P. Wen and published by John Wiley & Sons. This book was released on 2014-10-06 with total page 452 pages. Available in PDF, EPUB and Kindle. Book excerpt: Vaccine Manufacturing and Production is an invaluable reference on how to produce a vaccine - from beginning to end - addressing all classes of vaccines from a processing, production, and regulatory viewpoint. It will provide comprehensive information on the various fields involved in the production of vaccines, from fermentation, purification, formulation, to regulatory filing and facility designs. In recent years, there have been tremendous advances in all aspects of vaccine manufacturing. Improved technology and growth media have been developed for the production of cell culture with high cell density or fermentation. Vaccine Manufacturing and Production will serve as a reference on all aspects of vaccine production by providing an in-depth description of the available technologies for making different types of vaccines and the current thinking in facility designs and supply issues. This book will provide insight to the issues scientists face when producing a vaccine, the steps that are involved, and will serve as a reference tool regarding state-of-the-art vaccine manufacturing technologies and facility set-up. Highlights include: Comprehensive coverage of vaccine production : from a process point of view- fermentation to purification to formulation developments; from a production point of view - from facility design to manufacturing; and from a regulatory point of view - requirements from government agencies Authors from different major pharmaceutical and biotechnology companies Describes the challenges and issues involved in vaccine production and manufacturing of the different classes of vaccines, an area not covered by other books currently on the market
Download or read book Adenoviral Vectors for Gene Therapy written by David T. Curiel and published by Academic Press. This book was released on 2016-03-10 with total page 870 pages. Available in PDF, EPUB and Kindle. Book excerpt: Adenoviral Vectors for Gene Therapy, Second Edition provides detailed, comprehensive coverage of the gene delivery vehicles that are based on the adenovirus that is emerging as an important tool in gene therapy. These exciting new therapeutic agents have great potential for the treatment of disease, making gene therapy a fast-growing field for research. This book presents topics ranging from the basic biology of adenoviruses, through the construction and purification of adenoviral vectors, cutting-edge vectorology, and the use of adenoviral vectors in preclinical animal models, with final consideration of the regulatory issues surrounding human clinical gene therapy trials. This broad scope of information provides a solid overview of the field, allowing the reader to gain a complete understanding of the development and use of adenoviral vectors. Provides complete coverage of the basic biology of adenoviruses, as well as their construction, propagation, and purification of adenoviral vectors Introduces common strategies for the development of adenoviral vectors, along with cutting-edge methods for their improvement Demonstrates noninvasive imaging of adenovirus-mediated gene transfer Discusses utility of adenoviral vectors in animal disease models Considers Federal Drug Administration regulations for human clinical trials
Download or read book Bioreactor Design Concepts for Viral Vaccine Production written by Surajbhan Sevda and published by Elsevier. This book was released on 2024-05-12 with total page 476 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioreactor Design Concepts for Viral Vaccine Production covers a range of interdisciplinary chapters from the engineering perspective of bioreactor design to the biotechnological perspectives of vector design for vaccine development. The book covers bioreactor concepts such as static systems, single-use systems, stirred tanks, perfusion, wave and packed-beds. It reviews options for efficient and economical production of human vaccines and discusses basic factors relevant for viral antigen production in mammalian cells, avian cells, and insect cells. This book will be a great resource for those interested in implemented novel bioreactor design or experimental schemes towards intensified or/and enhanced vaccine production. Covers the fundamentals of bioreactor designs Provides strategies for designing a successful vector-based vaccine Discusses the applications of biological kinetics, thermodynamics and basic substrate requirements for viral vaccine production
Download or read book The Application of Viruses to Biotechnology written by Carla Varanda and published by Mdpi AG. This book was released on 2021-12-21 with total page 306 pages. Available in PDF, EPUB and Kindle. Book excerpt: Viruses are microscopic agents that exist worldwide and are present in humans, animals, plants, and other living organisms in which they can cause devastating diseases. However, the advances of biotechnology and next-generation sequencing technologies have accelerated novel virus discovery, identification, sequencing, and manipulation, showing that they present unique characteristics that place them as valuable tools for a wide variety of biotechnological applications. Many applications of viruses have been used for agricultural purposes, namely concerning plant breeding and plant protection. Nevertheless, it is interesting to mention that plants have also many advantages to be used in vaccine production, such as the low cost and low risks they entail, showing once more the versatility of the use of viruses in biotechnology. Although it will obviously never be ignored that viruses are responsible for devastating diseases, it is clear that the more they are studied, the more possibilities they offer to us. They are now on the front line of the most revolutionizing techniques in several fields, providing advances that would not be possible without their existence. In this book there are presented studies that demonstrate the work developed using viruses in biotechnology. These studies were brought by experts that focus on the development and applications of many viruses in several fields, such as agriculture, the pharmaceutical industry, and medicine.
Download or read book Synthetic DNA Delivery Systems written by Dan Luo and published by Springer Science & Business Media. This book was released on 2003-09-30 with total page 160 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA delivery into cells is a rapidly developing area in gene therapy and biotechnology. Moreover, it is a powerful research tool to determine gene structure, regulation, and function. Viral methods of DNA delivery are well-characterized and efficient, but little is known about the toxicity and immunogenecity of viral vectors. As a result, non-viral, transfection methods of DNA delivery are of increasing interest. Synthetic DNA Delivery Systems is a comprehensive and current resource on DNA transfection. The use of histidine-rich peptides and polypeptides as DNA delivery systems and self-assembled delivery systems based on cationic lipids and polymers are discussed. Targeted delivery to organelles, tumor cells and dendritic cells comprise an important topic.
Download or read book Development of Sustainable Bioprocesses written by Elmar Heinzle and published by John Wiley & Sons. This book was released on 2007-01-11 with total page 316 pages. Available in PDF, EPUB and Kindle. Book excerpt: Bioprocess technology involves the combination of living matter (whole organism or enzymes ) with nutrients under laboratory conditions to make a desired product within the pharmaceutical, food, cosmetics, biotechnology, fine chemicals and bulk chemicals sectors. Industry is under increasing pressure to develop new processes that are both environmentally friendly and cost-effective, and this can be achieved by taking a fresh look at process development; - namely by combining modern process modeling techniques with sustainability assessment methods. Development of Sustainable Bioprocesses: Modeling and Assessment describes methodologies and supporting case studies for the evolution and implementation of sustainable bioprocesses. Practical and industry-focused, the book begins with an introduction to the bioprocess industries and development procedures. Bioprocesses and bioproducts are then introduced, together with a description of the unit operations involved. Modeling procedures, a key feature of the book, are covered in chapter 3 prior to an overview of the key sustainability assessment methods in use (environmental, economic and societal). The second part of the book is devoted to case studies, which cover the development of bioprocesses in the pharmaceutical, food, fine chemicals, cosmetics and bulk chemicals industries. Some selected case studies include: citric acid, biopolymers, antibiotics, biopharmaceuticals. Supplementary material provides hands-on materials so that the techniques can be put into practice. These materials include a demo version of SuperPro Designer software (used in process engineering) and models of all featured case studies, excel sheets of assessment methods, Monte Carlo simulations and exercises. Previously available on CD-ROM, the supplementary material can now be accessed via http://booksupport.wiley.com by entering the author name, book title or isbn and clicking on the desired entry. This will then give a listing of all the content available for download. Please read any text files before downloading material.
Download or read book Lentiviral Vectors written by Didier Trono and published by Springer Science & Business Media. This book was released on 2001-12-14 with total page 276 pages. Available in PDF, EPUB and Kindle. Book excerpt: For the first time a compilation of chapters that depict the biological bases underlying the development of lentiviral vectors, the techniques involved in the manufacture of this new gene delivery tool, and its most promising applications.
Download or read book DNA Vaccines written by Mark W. Saltzman and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the early 1990s, almost 200 yr after Edward Jenner demonstrated the effectiveness of the smallpox vaccine, a new paradigm for vaccination emerged. The conventional method of vaccination required delivery of whole pathogens or structural subunits, but in this new approach, DNA or genetic information was administered to elicit an immunological response. Once it was observed that plasmid DNA delivered in vivo led to production of an encoded transgene (1), two ground-breaking studies demonstrated that immunological responses could be generated against antigenic transgenes via plasmid DNA delivered by DNA vaccination (as this approach is called) (2,3). The appe- ance of this new vaccination strategy coincided with advances in molecular biology, which provided new tools to study and manipulate the basic elements of an organism’s genome and also could also be applied to the design and production of DNA vaccines. DNA Vaccines is a major updated and enhancement of the first edition. It reviews state-of-the-art methods in DNA vaccine technology, with chapters describing DNA vaccine design, delivery systems, adjuvants, current appli- tions, methods of production, and quality control. Consistent with the approach of the Methods in Molecular Medicine series, these chapters contain detailed practical procedures on the latest DNA vaccine technology. The enthusiasm for DNA vaccine technology is made clear by the number of research studies published on this topic since the mid-1990s.
Download or read book Bioprocess Engineering Principles written by Pauline M. Doran and published by Elsevier. This book was released on 1995-04-03 with total page 455 pages. Available in PDF, EPUB and Kindle. Book excerpt: The emergence and refinement of techniques in molecular biology has changed our perceptions of medicine, agriculture and environmental management. Scientific breakthroughs in gene expression, protein engineering and cell fusion are being translated by a strengthening biotechnology industry into revolutionary new products and services. Many a student has been enticed by the promise of biotechnology and the excitement of being near the cutting edge of scientific advancement. However, graduates trained in molecular biology and cell manipulation soon realise that these techniques are only part of the picture. Reaping the full benefits of biotechnology requires manufacturing capability involving the large-scale processing of biological material. Increasingly, biotechnologists are being employed by companies to work in co-operation with chemical engineers to achieve pragmatic commercial goals. For many years aspects of biochemistry and molecular genetics have been included in chemical engineering curricula, yet there has been little attempt until recently to teach aspects of engineering applicable to process design to biotechnologists. This textbook is the first to present the principles of bioprocess engineering in a way that is accessible to biological scientists. Other texts on bioprocess engineering currently available assume that the reader already has engineering training. On the other hand, chemical engineering textbooks do not consider examples from bioprocessing, and are written almost exclusively with the petroleum and chemical industries in mind. This publication explains process analysis from an engineering point of view, but refers exclusively to the treatment of biological systems. Over 170 problems and worked examples encompass a wide range of applications, including recombinant cells, plant and animal cell cultures, immobilised catalysts as well as traditional fermentation systems. * * First book to present the principles of bioprocess engineering in a way that is accessible to biological scientists * Explains process analysis from an engineering point of view, but uses worked examples relating to biological systems * Comprehensive, single-authored * 170 problems and worked examples encompass a wide range of applications, involving recombinant plant and animal cell cultures, immobilized catalysts, and traditional fermentation systems * 13 chapters, organized according to engineering sub-disciplines, are groupled in four sections - Introduction, Material and Energy Balances, Physical Processes, and Reactions and Reactors * Each chapter includes a set of problems and exercises for the student, key references, and a list of suggestions for further reading * Includes useful appendices, detailing conversion factors, physical and chemical property data, steam tables, mathematical rules, and a list of symbols used * Suitable for course adoption - follows closely curricula used on most bioprocessing and process biotechnology courses at senior undergraduate and graduate levels.
Download or read book Bioprocess and Analytics Development for Virus based Advanced Therapeutics and Medicinal Products ATMPs written by Saurabh Gautam and published by Springer Nature. This book was released on 2023-08-10 with total page 448 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book reviews the knowledge, methods and available techniques in the rapidly advancing field of virus based vaccines and gene therapeutics. It also highlights new innovative tools and interdisciplinary techniques for bioprocess development and analytics of viruses and viral vectors. As such, it provides a timely and highly relevant resource, since current advances in pharmaceutical research have seen the rise of vaccines and advanced therapeutics and medicinal products (ATMPs), that rely on the power of viruses. However, developing bioprocesses and analytics required to create this often called “magic bullet” (i.e. gene therapy) remains an extremely challenging and costly task. This book offers strategies for overcoming hurdles and difficulties within in all the necessary steps of viral vector development - from scalability to purification methods and quality control. The book is intended for researchers working in academia or industry, as well as graduate students pursuing a career in virology.
Download or read book Non conventional Yeasts from Basic Research to Application written by Andriy Sibirny and published by Springer. This book was released on 2019-08-12 with total page 568 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume scopes several aspects of non-conventional yeast research prepared by the leading specialists in the field. An introduction on taxonomy and systematics enhances the reader’s knowledge on yeasts beyond established ones such as Saccharomyces cerevisiae. Biotechnological approaches that involve fungal utilization of unusual substrates, production of biofuels and useful chemicals as citric acid, glutathione or erythritol are discussed. Further, strategies for metabolic engineering based on knowledge on regulation of gene expression as well as sensing and signaling pathways are presented. The book targets researchers and advanced students working in Microbiology, Microbial Biotechnology and Biochemistry.
Download or read book Cell Culture Bioprocess Engineering Second Edition written by Wei-Shou Hu and published by CRC Press. This book was released on 2020-03-06 with total page 466 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is the culmination of three decades of accumulated experience in teaching biotechnology professionals. It distills the fundamental principles and essential knowledge of cell culture processes from across many different disciplines and presents them in a series of easy-to-follow, comprehensive chapters. Practicality, including technological advances and best practices, is emphasized. This second edition consists of major updates to all relevant topics contained within this work. The previous edition has been successfully used in training courses on cell culture bioprocessing over the past seven years. The format of the book is well-suited to fast-paced learning, such as is found in the intensive short course, since the key take-home messages are prominently highlighted in panels. The book is also well-suited to act as a reference guide for experienced industrial practitioners of mammalian cell cultivation for the production of biologics.
Download or read book Commercial Biotechnology written by and published by . This book was released on 1984 with total page 28 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Gene Vaccines written by Josef Thalhamer and published by Springer Science & Business Media. This book was released on 2011-08-31 with total page 332 pages. Available in PDF, EPUB and Kindle. Book excerpt: The induction of antigen-specific immune responses after in vivo transfection with expression plasmids has triggered a revolution of vaccine research. After a first hype, evoked by the fascinating options of this method, clinical studies did not reach the ambitious aims and a phase of disillusion ensued. It became obvious that Gene vaccines displayed a weaker immunogenicity in humans than had been observed in the mouse models. Meanwhile these hurdles have been overcome and gene vaccines undergo a renaissance. The present book gives an update of the “world of naked gene vaccines”, namely DNA and RNA vaccines. Its content ranges from general mechanisms, inherent immunostimulatory properties and the vast potential to modulate immune responses, to recent successful clinical studies and approved veterinary gene vaccines. Beyond the state-of-the-art of genetic immunization, the reader will be stimulated with a chapter addressing “burning questions”.
