Recombinant pharmaceutical manufacturing from plants - the future of molecular farming

www.regonline.com/farm10

9:00 – 9:45 Registration

9:45 – 10:00 Introduction by the Chair: Professor Ian A Graham, CNAP Director & Weston Chair of Biochemical Genetics, The University of York, UK

10:00 – 10:40 Development of a virus-derived system for the co-expression of multiple proteins at defined levels in plant cells

Dr. George Lomonossoff, John Innes Centre, UK

A major challenge in the development of plants as bioareactors has been to a devise system that can direct the synthesis of multiple proteins within the same cell at defined and differing levels. Such a system would enable protein complexes containing a variety of polypeptides in different amounts to be produced. It would also enable metabolic pathways which require different amounts of enzyme for each step to be constructed. This talk will concern the development of such a system which is based on sequences derived from Cowpea mosaic virus (CPMV). Though orginally intended for transient expression, the system has recently been modified for use in stable transformation, increasing its utility.

10:40 – 11:20 Increasing yields of medicinals by plant molecular breeding

Professor Ian A Graham, CNAP Director & Weston Chair of Biochemical Genetics, The University of York, UK

Modern molecular breeding methods will be introduced and used to demonstrate the new timelines that are now possible for the rapid improvement of medicinal plants as robust production systems

11:20- 11:25 Speakers photo

11:25 – 12:00 Mid-morning break

12:00 – 12:40 Tackling chronic disease through improvements to foods

Professor Cathie Martin, Norwich Research Park, UK

A major challenge for society is to reduce the frequency of the chronic diseases; cardiovascular disease, cancer and age-related degenerative diseases. Epidemiological studies have demonstrated the efficacy of diets high in fruit and vegetables in reducing the incidence of chronic disease because they contribute important phytonutrients which serve to promote antioxidant defence mechanisms. Plant biotechnology can make a very significant contribution to dietary improvement through model foods that test the importance of specific bioactives in promoting particular aspects of health, markers that allow molecular breeding for enhanced levels of bioactives and genetic engineering that provides novel, health-promoting (functional) foods.

12:40– 12:50 Expression of human papillomavirus l1 capsomeres in tobacco chloroplasts: a step towards cost-effective second-generation vaccines

Andreas Lössl University of Natural Resources and Applied Life Sciences, Austria

Various types of human papillomaviruses (HPV) are causatively associated with cervical carcinoma which is the second most common cancer in women worldwide. Most of the cervical cancer cases occur in developing countries. Due to limitations in the availability of currently used virus-like particle (VLP)-based vaccines against HPV to women of these countries, the development of a cost-effective second-generation vaccine is a necessity. Capsomeres have recently been demonstrated to be highly immunogenic and to have a number of advantages as a potential cost-effective alternative to VLP-based HPV vaccines. We have expressed a modified HPV-16 L1 (L1_2xCysM) gene that retained the ability to assemble L1 protein to capsomeres in tobacco chloroplasts. The recombinant protein yielded up to 1.5% of total soluble protein. Assembly of capsomeres was examined and verified by cesium chloride density gradient centrifugation and sucrose sedimentation analysis. An antigen capture enzyme-linked immunosorbent assay confirmed the formation of capsomeres by using a conformation specific monoclonal antibody which recognized the conformational epitopes. Transplastomic tobacco plants exhibited normal growth and morphology, but all such lines showed male sterility in the T₀, T₁ and T₂ generations. Taken together, these results indicate the possibility of producing a low-cost capsomere-based vaccine by plastids

12:50 – 13:40 Lunch and Poster Viewing

13:40 – 14:30 Discussion panel session

Please submit questions to Euroscicon staff during the event. These questions will be asked to the panel of speakers at this panel session. Plus you are free to ask additional questions during the session

14:30 – 15:10 Using thin-layer chromatography (TLC) to detect biologically-active compounds in plant extracts

Professor Peter Houghton, Emeritus Professor Pharmaceutical sciences Division, Kings College London

In the last 20 years bioassay-guided isolation of active compounds from plant extracts has been widely used in the discovery of new lead compounds. One disadvantage of the conventional method of testing each fraction is the large number of fractions that have to be tested. The use of TLC together with various spray reagents for activity of biological interest enables the early rejection of fractions showing no activity and also helps to quickly identify the type of phytochemical and its rapid isolation.

15:10 – 15:40 Afternoon Tea/Coffee and Last Poster Viewing

15:40 – 16:20 Containment strategies in biopharming

Professor Denis J Murphy, University of Glamorgan, UK

This paper examines the challenges of segregating biopharmed crops from mainstream crops, particularly those destined for food or feed use. One commercially viable strategy to limit or avoid biopharming intrusion into the human food chain is the rigorous segregation of food and non-food varieties of the same crop species via a range of either physical or biological methods. Even more secure segregation is possible by use of non-food crops, non-crop plants, or in vitro plant cultures as production platforms for biopharming. Such platforms already under development range from outdoor-grown Nicotiana spp. to glasshouse-grown Arabidopsis, lotus and moss. Even more effective methods for secure biocontainment include plastid expression of transgenes, inducible and transient expression systems, and physical containment of plants or cell cultures. In the current atmosphere of heightened concerns over food safely and biosecurity, the future of biopharming may be largely determined by the extent to which the sector is able to maintain public confidence via a more considered approach to containment and security of its plant production systems.

16:20 - 17:00 Metabolic engineering of high-value and nutritional isoprenoids in plants

Dr Paul Frazer, Royal Holloway University London, UK

Over the past decade genetic/metabolic engineering of isoprenoid biosynthesis and accumulation has resulted in the generation of transgenic varieties containing enhanced or altered isoprenoids. In achieving this important goal many fundamental lessons have been learnt. Most notably is the observation that the endogenous pathways in higher plants appear to resist engineered changes. Typically, this resistance manifests itself through intrinsic regulatory mechanisms that are “silent” until manipulation of the pathway is initiated. In the present presentation the progress made in the genetic engineering of isoprenoids in tomato fruit and other Solanaceae will be reviewed.

17:00 Chairman’s summing up.

This meeting was organised by Euroscicon (www.euroscicon.com), a team of dedicated professionals working for the continuous improvement of technical knowledge transfer to all scientists. Euroscicon believe that they can make a positive difference to the quality of science by providing cutting edge information on new technological advancements to the scientific community. This is provided via our exceptional services to individual scientists, research institutions and industry. The event was hosted by ‘BioPark (www.biopark.co.uk), a research and development centre in Welwyn Garden City providing specialist facilities and support for bioscience and health technology businesses to grow, and to develop new products and technologies

About the Chair

Ian A Graham, graduated from the Queen’s University of Belfast with a first class honours degree in Botany and Genetics in 1986. He obtained his PhD in Plant Molecular Biology from the University of Edinburgh in 1989, after which he went on to do postdoctoral research at the University of Oxford and The Carnegie Institution Plant Biology Laboratory at Stanford University. He took up his first faculty position at The University of Glasgow in 1993 and moved to the University of York in 1999 where he holds the Weston Chair of Biochemical Genetics and is current Director of CNAP (http://www.york.ac.uk/org/cnap/)

About the Speakers

Denis J Murphy; PhD at Univ of York, UK; Fulbright postdoc fellowship at Univ California Davis; Royal Society postdoc fellowship at Australian National Univ, Canberra; Lecturer in Molecular Biology at Univ Durham; Head of Oilseeds Research Dept at John Innes Centre UK. Currently Head of Biotechnology Unit at University of Glamorgan, UK. Also Biotechnology Advisor to United Nations Food and Agriculture Organization and Chair, Biology Advisory Committee, Malaysian Palm Oil Board.

Professor Houghton retired as Professor in Pharmacognosy at KCL in 2008, after over 30 years working there.

He has published over 250 research papers on the chemistry and biological activity of plants and their constituents. His research areas include substances from plants potentially useful in wound healing, cancer and neurodegenerative disease.

Cathie Martin is a group leader at the John Innes Centre, the leading plant research institute in Europe. She is Professor at the University of East Anglia and Niels Bohr Professor in the Faculty of Life Science, University of Copenhagen, Denmark. Recently she has been co-ordinating research into how diet can help to maintain health and reduce the risk of chronic disease, and how crops can be fortified to improve diets. Her interests span from fundamental to applied plant science. She is Editor-in-Chief of The Plant Cell, and the first woman and the first non-American to hold this post.

Paul Frazer has over 20 year’s experience working both in academia and industry within the UK and abroad. During this period Dr Fraser has worked on the analysis, biosynthesis, regulation and metabolic engineering of carotenoids and isoprenoids, both in plant and microbial systems.

Andreas Lössl graduated at the University Giessen in 1991. He worked at the Universities of Munich (TUM: 1995-2000) and LMU (2000-2004). From 2004 to 2010 he developed a research group at the University of Natural Resources and Life Sciences, Vienna. He is focussed on plant organellar genomes and their value for biotechnological use. His group was the first who managed:

^1. Synthesis of polyesters in chloroplasts transformed with a bacterial 3-gene operon.

^2. Regulation of chloroplast-harboured transgene expression by an inducible T7-RNA polymerase.

^3. Expression of novel antigen-vaccines against HPV virus in chloroplasts

George Lomonossoff graduated from the University of Cambridge in 1976. His Ph.D. studies, at MRC Laboratory of Molecular Biology (LMB), Cambridge concerned the assembly of Tobacco mosaic virus and were followed by an MRC Post-doctoral fellowship at LMB Cambridge to determine sequence of TMV RNA. He moved to the John Innes Centre Norwich in 1980 and has continued to work there since apart from two periods: as a Fulbright scholar, at Cornell University, USA (1987-1988) and as a Visiting Researcher, at The Scripps Research Institute, La Jolla, USA, (1998). George’s research has focused on the molecular biology of RNA plant viruses and their use for the production of foreign proteins in plants . He is an honorary professor and UEA and has co-ordinated several EU FP consortia concerned with the production of plant-derived vaccines.

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