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

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.