International Conference and Exhibition on Antibodies August 10-11, 2015 Birmingham, UK

Biography:

Mike is a co-founder of IONTAS Ltd, a therapeutic antibody discovery company. He constructed the IONTAS antibody phage display library, is a co-inventor of the IONTAS mammalian cell antibody and T cell receptor display platform and acts as the project leader for several therapeutic antibody discovery projects. Mike was previously Head of Protein Engineering at Acambis plc (acquired by Sanofi) and was the Project Leader, responsible for protein expression, within the Atlas Project at the Wellcome Trust Sanger Institute. He has over 20 years experience in cell and molecular biology gained during research at the Massachusetts Institute of Technology, Universities of Edinburgh and Cambridge and the biotechnology industry. He has published 30 papers and book chapters (cited over 1000 times, h-index 17) and is inventor on 5 US patent and patent applications.

Abstract:

The availability of large libraries of binders expressed within mammalian cells will allow direct screening for function through simultaneous expression and functional reporting within the same cell. In addition, libraries of binders in mammalian cells could also be used for display of binders on the cell surface allowing the screening of millions of clones by flow sorting while providing information on both the level of expression and the extent of binding within individual clones. The main limitation in achieving this has been the inability to construct sufficiently large libraries containing a single antibody gene/cell. We have solved this problem by directing the integration of antibody genes into a single genomic locus through the use of site-specific nucleases. Antibody libraries consisting of many millions of clones have been constructed and binders selected. This includes antibodies formatted as IgGs expressed in the cell type used for production.

Biography:

Jan Voskuil is the Chief Scientific Officer at antibody manufacturer Everest Biotech in Oxfordshire, UK. After specializing in prokaryotic cell biology through his PhD program in Amsterdam, Netherlands, and a Postdoc position at Stanford, California, he switched to the science of neurodegenerative diseases at Oxford, UK through Postdoc positions at Dunn School of Pathology and at MRC and through a leading position at the Alzheimer drug discovery company Synaptica. He subsequently gained experience in a GLP-regulatory environment in CRO companies both in Oxfordshire and Cambridgeshire, validating assays in Flow Cytometry and ELISA platforms and writing SOPs. His extensive experience with generating and characterizing monoclonal and polyclonal antibodies in combination with accrued knowledge on most immunoassays in academic and commercial environments made him the ideal candidate to take charge in putting Everest Biotech on the global map by ever raising the quality and size of its catalogue and by delivery of adequate technical support. As a result, Everest antibodies are currently part of most globally well-known catalogues, and its products are increasingly recognized as useful alternatives to unfit monoclonal antibodies.

Abstract:

Antibodies play a pivotal role in diagnostics, biomarkers research and increasingly in therapeutics. However in all three fields problems related to specificity and to consistency of the antibodies occur. Such problems are being kept to a minimum by the use of monoclonal antibodies, because of their immortality and for their single molecular basis. However, existing monoclonal antibodies have generally not been screened for the specific application it is currently required for. This leads to the use of antibodies with insufficient affinity for that particular application. In addition different formulations at which one antibody is offered to the market will cause inconsistencies in performance. Any polyclonal antibody is dismissed because it represents a mixture of different molecules, and therefore this mixture will change in composition from animal to animal thus changing the characteristics of the reagent from batch to batch. Although for therapeutics the best way to address these problems may be through the route of recombinant reagents (avoiding the animals altogether), one remains to be vigilant for cross-reactivity of reagents when reacting to a non-unique epitope. Here is a good opportunity to make the most of a cost-effective alternative for in vitro and for ex vivo tests; there is evidence that epitope-specific polyclonal antibodies offer an adequate alternative to unavailable fit-for-purpose monoclonal antibody.

Biography:

Hiroshi Ueda has completed his PhD in the University of Tokyo, and studied antibody engineering in Laboratory of Molecular Biology, Medical Research Council, Cambridge, UK. He has been working with antibodies with the aim of developing novel methods for biosensing and diagnostics for years, and received many awards including Biosensors 2012 Best Paper Award. He is now the Vice chairman of Biochemical Assay Society of Japan, and a Councilor of The Japanese Biochemical Society.

Abstract:

In the areas of biological analysis and clinical diagnostics, rapid and specific quantification for various molecules is considered highly important. Here we describe a novel reagent less fluorescent biosensor strategy based on the antigen-dependent removal of a quenching effect on fluorophores attached to an antibody. Using position-specific protein labeling systems, we found that an antibody single chain variable region (scFv) fluoro labeled at the N-terminal region showed a significant antigen-dependent fluorescence increase. Investigation of the enhancement mechanism by mutagenesis, denaturation, and fluorescence lifetime measurement of the carboxytetramethylrhodamine (TAMRA)-labeled anti-osteocalcin scFv showed that antigen-dependency was dependent on semi-conserved tryptophan residues. This suggested that the binding of the antigen led to the removal of quenching effect caused by the proximity of tryptophan residues to the linker-tagged fluorophore. Using TAMRA-scFv, many targets including peptides, proteins, and haptens including narcotics could be quantified. Similar or higher sensitivities to those observed in competitive ELISA were obtained, even in human plasma. Furthermore, we found that a number of Fab fragments fluorescence-labeled at their N-termini with various dyes showed higher fluorescence enhancements probably due to enhanced quenching. Due to its versatility, these “quenchbodies” are expected to have a range of applications, from in vitro diagnostics to imaging of various targets in situ. Latest results for application to cellular imaging will be shown.

Biography:

Hazel Gorham, PhD, Director, Biosimilars Development, Scientific Affairs, has 26 years of experience in both academic and pharmaceutical research/industry across a wide range of roles including developing Clinical Development Plans for biosimilars. Dr. Gorham has experience in all aspects of biosimilar development including study design and regulatory agency discussions. She has experience in all aspects of clinical trial project management and has managed the conduct of multiple Phase I through IV clinical trials and post-marketing studies across geographical regions and differing therapeutic areas. Dr Gorham completed her post-doctoral graduate studies in microbiology at the University of Warwick, UK.

Abstract:

The market of biologics is growing at nearly twice the rate of pharma as a whole. The expiration of patents and other intellectual property rights for originator biologicals over the next decade opens up opportunities for biosimilars to enter the market and increase industry competition. In order to be cost effective a biosimilar product needs access to global markets based on a single development programme that meets the requirement of regulators internationally. Despite increasing alignment in the regulatory requirements for biosimilars between EMA, FDA, WHO and other jurisdiction, there are still many scientific and practical challenges for demonstrating biosimilarity and interchangeability including scientific factors, drug interchangeability and statistical considerations.

Biography:

Pisitkun Trairak is currently working as Principal Investigator, Chulalongkorn University Systems Biology (CUSB) Center, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand. He has successfully completed his Administrative responsibilities as Senior Scientist, InterPrET Research Center, Department of Biomedicine, Aarhus University, Denmark. His research has included Systems Biology, Computer Software and Web Application Development. Based on this research and fellowship training, he has received several awards and honors, such as Outstanding Mentorship Award, National Heart, Lung, and Blood Institute, National Institutes of Health.

Abstract:

In the current era of large-scale biology, systems biology has evolved as a powerful approach to identify complex interactions within biological systems. In addition to high throughput identification and quantification techniques, methods based on high-quality mono-specific antibodies remain an essential element of the approach. To assist the large-scale design and production of peptide-directed antibodies for systems biology studies, we developed a fully integrated online application, AbDesigner to help researchers select optimal peptide immunogens for antibody generation against relatively disordered regions of target proteins. Here we describe AbDesigner in terms of its features, comparing it to other software tools, and the applications of it to design various antibodies.

Biography:

Saroj Velamakanni earned his PhD degree in Molecular Microbiology at University of Cambridge, UK. In 2002, he was awarded the prestigious Cambridge Nehru Scholarship to pursue his Doctoral studies. After Post-doctoral work at the Universities of Cambridge, he received a research fellowship from Wolfson College, Cambridge to work on Calcium Signalling in mammalian systems. During this time his work was published in prestigious journal Nature. In 2012, he became associated with the start-up Biotech/in vitro diagnostics company Fahy Gurteen Labs based in Cambridge Research Park first as a Senior R&D Scientist and currently as a Research Director. He has 22 peer reviewed high impact publications and six patents to his credit.

Abstract:

Cell cycle related molecules play a pivotal role in maintaining genomic stability and can be used as biomarkers for cell cycle phase distribution. In this study, we performed immunohistochemical multiparameter analysis using five cell cycle related biomarkers for a cohort of 182 patients and linked it with clinicopathological information. Various stages involved in the design, development and validation of proprietary antibodies against these biomarkers will be discussed. Three unique cell cycle phenotypes were identified 1) out-of-cycle state 2) G1 delayed/arrested state and 3) accelerated cell cycle progression state. Our algorithm sheds new light into the cell cycle state of dynamic tumour populations in vivo. The information obtained is of major prognostic significance and may impact on individualised therapeutic decisions.

Biography:

Reem Hamdy A Mohammed is a Assistant Professor of Rheumatology, graduated from the School of Medicine, Cairo University, MB, BSCh, 1995 excellent grades with honors. She finished her Master degree of Rheumatology and Rehabilitation in May 2000, and MD Rheumatology and Rehabilitation, PhD rheumatology, at Cairo University in 2004. She is recognized as international fellow by the American college of Rheumatology in 2011, nominated as Fellow by the Royal College of Physicians UK in May, 2014 and member of the American Association of Immunologists. As a rheumatologist, she is committed to disseminating evidence based knowledge in the field of rheumatology and autoimmune diseases to all her students, advancing research and academic standards in my organization. In the field of research, she shared in multiple researches in the field of autoimmune diseases many of which has been published in addition to sharing as an author in two book publications about vasculitis. She is also serving as an editor and reviewer in a number of Journals in the field of Rheumatology and Clinical Immunology.

Abstract:

With the evolution of biologic drugs in the last one and half decades, the treatment paradigm of autoimmune diseases experienced a dramatic shift that ultimately improved outcome in the setting of refractory illness. Targeting the antibody producing B lymphocyte at its different developmental stages represents one important approach nowadays in the treatment strategy of a multiplicity of autoimmune diseases especially rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). The production and differentiation of the B cells passes through various developmental stages with the earliest identifiable precursor being the stem cells also known as the progenitor B-Cell, then pro-B, pre-B, immature B, virgin B-Cell and plasma cells. Therapeutic antibodies to cell surface receptors on B lymphocytes as well as their survival factors have been extensively tested with the anti-CD-20 therapy Rituximab being on top of the list used for refractory rheumatoid and in refractory systemic lupus particularly after failure of conventional immunosuppressive drugs, in addition to anti B lymphocyte survival factor anti- BLys like Atacicept and Belimumab, the recently approved biologic therapy for systemic lupus erythematosus. Other uprising B cell targeted strategies include Blisimod (A623) soluble and membrane bound BLyS, and Tabalumab (LY2127399) soluble and membrane BLyS, such novel drugs are being currently evaluated in SLE and RA

Biography:

Dr Zhang-lijuan (MD, phD ) is the Director of the Dept. of Rickettisology, National Institute for Communicable Disease Control and Prevention, China CDC. She is and PhD Candidate Supervisor and one of the members of the Committee of Natural Focus Disease, Ministry of Hygiene of People’s Republic of China, and a member of the National Zoonoses Committee. Her research interests are national surveillance of Rickettsioses, Anaplasmosis and Ehrlichiosis and molecular epidemiological and pathogenesis researches of rickettsiae, anaplasma and ehrlichia .She has published more than 60 papers on the surveillance of rickettsiae and basic research on rickettsiae agents.

Abstract:

More and more epidemiological evidences indicated that the annual numbers of the emerging anapalsema and ehrlichia and the reemerging rickettsiae have steadily increased in China and the outbreaks of these zoonoses have been frequently reported in recent years. Of the traditional rickettsioses, scrub typhus, caused by Orientia tsutsugamushi (Ot), is one of the serious acute febrile illness in China. In some severe endemic areas of southern China, hundreds of farmers suffered scrub typhus in small villages each year. Similarly, murine typhus, caused by R. typhi is highly prevalent in some rural areas although the louse borne epidemiological typhus has been controlled in China. Noticeably, a big outbreak of murine typhus involved 76 patients occurred in drug detoxification program from Ruili city, Yunnan province in 2010. Besides R. sibirica and R. heilongjiangensis, emerging spotted fever group rickettsiae (SFGR) including R. raoulti, R. felis, R. massiliae and SFGR Hainan-1 were successively detected in China in recent years. Specifically, a highly seroprevalence of the SFGR Hainan-1 were identified in 46.1% of the adult population and 37.5% of the preschool children in the local areas. An active national investigation indicated that the emerging tick born E. chaffeensis and A. phagocytophilum were highly prevalent among agrarian residents, domestic animals and some wild animals. In particular, an unusual transmission of nosocomial cases of human granulocytic anaplasmosis occurred in Anhui Province in 2006. In China, the rickettsial diseases are the greatest challenge to public health. Notably, rural residents are not only at increased risk of these zoonoses but also these diseases are mostly under-recognized.

Biography:

William G Whitford is Sr. Manager, Cell Culture, GE Healthcare in Logan, UT with over 20 years experience in biotechnology product and process development. He joined the company 12 years ago as a team leader in R&D developing products supporting biomass expansion, protein expression and virus secretion in mammalian and invertebrate cell lines. Products he has commercialized include, defined and animal product-free hybridoma media, fed-batch supplements, and aqueous lipid dispersions. An invited lecturer at international conferences, he has published over 250 articles, book chapters and patents in a number of fields in the bioproduction arena. He now enjoys such industry activities as serving on the editorial advisory board for BioProcess International.

Abstract:

Upstream continuous biomanufacturing is supported by a number of new approaches to perfusion culture. This includes continuing developments in single-use systems and cell-culture media design. SU perfusion-capable systems support a high degree of flexibility in process flow and configuration, component types, physical location and production mode. Continuous biomanufacturing through bioproduction in a high-cell density perfusion culture is an exciting potential. Data from a structured approach to developing a perfusion medium from a combination of commercially available medium and feeds will be presented. Insights as to how this methodology can be applied to other cell culture media will be outlined. Optimization of culture media and process fluids for such intensified processes also includes such new ideas as heightened process understanding, the relative value of powder vs. bulk liquid and in-line fluids conditioning. Employment of such is enabled by higher specific productivities, regional manufacturing and improved shipping/packaging technologies. These activities support the ultimate goal of real-time, continuous quality and process verification in pre-engineered, modular multi-product manufacturing facilities.

Biography:

Dr. Ming Yang is an immunologist at the National center for foreign animal diseases/Canadian food inspection agency, where she produces and characterizes monoclonal antibodies against vesicular disease viruses such as foot-and-mouth disease (FMD) virus, vesicular stomatitis (VS) virus and swine vesicular disease (SVD) virus. She developed several immune assays for the diagnoses of vesicular diseases, such as ELISA and lateral flow strip tests. Dr. Yang published more than 20 manuscripts in the peer-reviewed journals. She graduated from University of Manitoba with M.Sc. degree in immunology in 1992 and PhD. in Microbiology in 2000.

Abstract:

Foot-and-mouth disease (FMD) is one of the most highly contagious diseases that affects cloven-hoofed animals such as cattle, pigs, sheep and goats. FMD is an economically devastating disease that severely constrains the international trade of animals. FMD virus (FMDV) has seven serotypes: O, A, C, Asia 1, SAT 1, SAT 2 and SAT 3based on serological tests. Among the seven serotypes of FMDV, O and A are the most widespread and currently found in Africa, the Middle East, Asia, limited area of South America and sporadically in Europe. The objective of this study was to generate and characterize monoclonal antibodies against foot-and-mouth disease virus serotype A. Using the produced mAbs develop a reliable and rapid competitive ELISA (cELISA) for the detection of antibodies to FMDV serotype A. A panel of FMDV/A specific mAbs were generated from this study. The binding epitopes of the mAbs were characterized. Two of the twelve mAbs’ biding sites are located on VP2 and VP3 as previously identified antigenic site 3 using the monoclonal antibody resistant mutant selection method. These two mAbs clearly inhibited the binding of a FMDV/A polyclonal serum to FMD serotype A viruses. Thus acELISA for FMDV serotype A antibody detection was developed using the two serotype A-specific mAbs and the inactivated virus as the antigen. The cut-off value was set at <50% of inhibition based on the tested results of a total of 1,174 negative sera. Three of the 1,174 negative samples exceeded this cut-off value, which produced a diagnostic specificity of 99.7%. The antibody responses to FMDV/A in experimentally inoculated animals were examined using the A/cELISA. All samples demonstrated positive antibody responses starting at 5 days post inoculation (dpi) and remained positive until the end of the experiment (21-28 dpi). The results showed that the A/cELISA detected antibodies against FMDV/A in all tested animal species (cattle, pig and sheep). Serum samples from vaccinated (A22 Iraq) and challenged (A/Vietnam/13) sheep were examined using the A/cELISA and compared with the virus neutralization test (VNT) recognized by the OIE as a standard method. All sheep (100%) demonstrated a positive seroconversion at 10 dpc using A/cELISA and 60% of the sheep showed positive results using a VNT. The results showed that the performance of this A/cELISA was comparable or better than the VNT indicating the potential of this cELISA as an alternative assay to VNT. This A/cELISA is a simple, reliable test to detect antibodies against FMD serotype A viruses. The test will be a useful tool for surveillance, epidemiological study of FMD and monitoring immune responses following FMDV/A vaccination.

Biography:

Abstract:

Background: Today, HIV infection is referred to as a serious threat to humanity. Many efforts have been made to prepare an effective vaccine, but have remained inconclusive. Therefore, new strategies in order to increase the immune response to vaccines using immunological adjuvant such as TLR agonists like the Pseudomonas aeruginosa flagellin (FliC) were presented. In this study, given the effect of immunization route on the immune response induced by vaccination, candidate vaccine HIV-1p24-Nef conjugated to FliC molecule was injected in different routes and the immune responses were evaluated. Materials & Methods: BALB/c mice were distributed into 6 groups and immunized with 20 μg/100μl of HIV-1 p24-Nef conjugated to FliC, p24-Nef and FliC which were prepared in Montanide ISA70, subcutaneously and intramuscular three times under the same conditions. Two weeks after the final boosting, lymphocyte proliferation was measured by Brdu method, the response of IL-4 and IFN-γ cytokines, as well as the level of total antibodies and their isotypes were evaluated using ELISA method. Also IFN-γ ELISPOT was performed to detect the memory T cells frequency. Results: Results show that, in comparison with control groups, the conjugated HIV-1p24-Nef-FliC significantly increased lymphocyte proliferation responses, higher levels of cytokines responses and IFN-γ producing lymphocytes subcutaneously but the level of humoral immune responses significantly increased in the intramuscular route. Conclusion: FliC molecule could be used as adjuvant in combination with vaccines candidate against HIV-1.

Biography:

Ali Jazayeri, PhD, is head of protein engineering at Heptares Therapeutics. He received his undergraduate degree in generics from University of Manchester in 1999 and his PhD in Molecular Biology from Cambridge University in 2003. Ali joined Heptares in 2007 as the company was founded and led the technology development and industrialization of the receptor stabilisation methodology. In his current position, he is responsible for the protein engineering group that generates stabilized receptors for crystallisation and antibody generation. Prior to joining Heptares, he was a post-doctoral scientist at Cancer research UK Clare Hall laboratories and KuDOS pharmaceuticals working on DNA damage response pathways.

Abstract:

GPCRs represent excellent antibody targets given their central role in the pathology of many diseases and cell surface location. However GPCRs make poor antigens due to their conformational flexibility, low expression levels, inherent instability and hydrophobic nature. Using protein engineering approaches we create conformationally stabilised receptors (StaRs) that can be purified to high purity and homogeneity with enhanced half-life. StaRs allow generation of high quality antigens that can be used to raise functional antibodies.

Biography:

Dr Hou has completed his PhD at the age of 30 years from Birmingham University and postdoctoral studies from University College London School of Medicine. He has published more than 15 papers in reputed journals.

Abstract:

Common variable immunodeficiency (CVID) is a heterogeneous group of immunodeficiencies characterised by low immunoglobulin levels and recurrent infections, frequently accompanied by autoimmune complications. CTLA-4 is an essential immune regulator, which controls T cell activation and plays a critical role in Treg suppression. CTLA-4 acts by controlling the levels of CD28 signalling in T cells via competition for their shared ligands CD80 and CD86 on APCs. A major effect of this competition is to prevent the activation of self-reactive T cells and disruption leads to fatal autoimmunity in mice. Using whole exome and targeted sequencing approaches we have identified a number of individuals with heterozygous mutations in CTLA-4, including stop codon, splice-site and missense mutations. These mutations, albeit incompletely penetrant, were found in CVID patients and family members associated with severe autoimmunity. Characterisation of T cells from several patients as well as unaffected carriers revealed reduced CTLA-4 expression, which was clearly evident in CD4+Foxp3+ Treg. Two independent point mutations in the CTLA-4 extracellular domain were found to significantly reduce the ability to bind ligand and carry out transendocytosis. Consistent with murine data, individuals with CTLA-4 insufficiency were found to have expanded numbers of Foxp3-expressing Treg in their blood irrespective of disease status, which most likely reflects enhanced CD28 signalling in Treg. Nonetheless, Treg were functionally impaired when tested in suppression assays. Together, these data reveal that CTLA-4 insufficiency in humans is associated with altered Treg homeostasis and function, autoimmune dysregulation as well as B cell- and antibody deficits.