Day 2 :
Keynote Forum
David Meininger
TRIANNI, USA
Keynote: The Trianni Mouse — the next generation transgenic platform for the isolation of fully human monoclonal antibodies
Time : 09:05-09:40
Biography:
Dr. Meininger was most recently Senior Vice President, Corporate Development with OncoSec Medical after serving as Executive Director, Business Development & Licensing with Merck where he had global responsibility for biologics technologies as a member of the West Coast Innovation Hub. He previously managed biologic lead generation at Merck where his team progressed several immuno-oncology and other programs to clinical development. He also contributed as a core team member on multiple programs from discovery through the clinic at Amgen after completing his postdoc in Protein Engineering at Genentech. Dr. Meininger received his PhD from UCSD and his MBA from The University of Washington.
Abstract:
Trianni offers a fully humanized monoclonal antibody platform. The Trianni’s technology is distinguished by key advantages of ease of use, comprehensive antibody diversity and in vivo selection. Founded in 2010, The Trianni Mouse is an antibody discovery platform with several distinctive and highly attractive features. Chief among these is the fact that it expresses a full repertoire of variable domains for human antibody heavy and light chains. In generating its platform, Trianni employed a unique combination of in silico gene design, DNA synthesis and targeted genomic mutagenesis to modify all three mouse antibody chain loci (heavy, kappa and lambda). The new gene segments embedded in these loci have a novel proprietary optimized structure involving human coding sequences (of V, D and J) and mouse regulatory sequences (promoters, enhancers, introns, recombination signal sequences). This feature distinguishes the Trianni Mouse from all other similar platforms now available. The platform has been extensively validated both internally and externally (adopted by several major pharma companies) and has proven itself as a powerful resource for the isolation of high quality therapeutic monoclonal antibodies, i.e., effective in terms of hybridoma frequency, antibody titers, affinities and biological effector function. Trianni continues to enhance and expand the platform, generating mice that respond to evolutionary conserved sequences; respond to antigens with non-immunoglobulin scaffolds and produce bispecific antibodies.
Keynote Forum
William George Whitford,
GE Health Care, USA
Keynote: Round Table on Upstream Continuous Manufacturing Equipment and Materials
Time : 09:40-10:40
Biography:
William George Whitford is a Strategic Solutions Leader for BioProcess at GE Healthcare Life Sciences in Logan, UT with over 20 years experience in biotechnology product and process development. He has joined the company as an R&D Team Leader developing products supporting biomass expansion, protein expression and virus secretion in mammalian and invertebrate cell lines. He has published over 300 articles, book chapters and patents in a number of fields in the biotechnology arena.
Abstract:
Upstream continuous bioprocessing (CB) is supported by number of approaches to perfusion culture. Coincident with a growing interest in CB are continuing developments in single-use (SU) processing and cell-culture media design. SU perfusion-capable equipment is now available to support many types of process flexibilities, increased facility utilization as well as reduced CapEx, OpEx and build times. Such advanced SU systems support a high degree of flexibility in process flow and configuration, component types, physical location and production mode. As animal-cell platforms are adapted to perfusion culture in such facilities, the need for media and feeds optimized for intensified processes involving perfusion in SU equipment has become apparent. Renewed discussions appearing on the relative value of powder vs bulk liquid supply are based upon perfusion-based operation, higher specific productivities, regional manufacturing and improved shipping/packaging technologies. Formulation goals are being achieved through the implementation of such new development tools as HTS, ‘omics capabilities, chemometrics and metabolic flux analysis. The above activities support the ultimate goal of real-time, continuous quality and process verification in pre-engineered, modular and turn-key multi-product manufacturing facilities. For many platforms, such designs can also be imagined in either ready-to-use, microenvironment-based flexible factories or pre-assembled streamlined suite pods promoting closed processing within open-production halls or unclassified “ballroom†or “dance-floor†CNC suites. Such facilities might possess advanced in-line testing technologies, eventually establishing a global, enterprise process control integrating even the scheduling and management of such activities as raw material sourcing and equipment calibration.
Keynote Forum
William George Whitford
GE Health Care, USA
Keynote: Round Table on Upstream Continuous Manufacturing Equipment and Materials
Time : 09:40-10:40
Biography:
William George Whitford is a Strategic Solutions Leader for BioProcess at GE Healthcare Life Sciences in Logan, UT with over 20 years experience in biotechnology product and process development. He has joined the company as an R&D Team Leader developing products supporting biomass expansion, protein expression and virus secretion in mammalian and invertebrate cell lines. He has published over 300 articles, book chapters and patents in a number of fields in the biotechnology arena.
Abstract:
Upstream continuous bioprocessing (CB) is supported by number of approaches to perfusion culture. Coincident with a growing interest in CB are continuing developments in single-use (SU) processing and cell-culture media design. SU perfusion-capable equipment is now available to support many types of process flexibilities, increased facility utilization as well as reduced CapEx, OpEx and build times. Such advanced SU systems support a high degree of flexibility in process flow and configuration, component types, physical location and production mode. As animal-cell platforms are adapted to perfusion culture in such facilities, the need for media and feeds optimized for intensified processes involving perfusion in SU equipment has become apparent. Renewed discussions appearing on the relative value of powder vs bulk liquid supply are based upon perfusion-based operation, higher specific productivities, regional manufacturing and improved shipping/packaging technologies. Formulation goals are being achieved through the implementation of such new development tools as HTS, ‘omics capabilities, chemometrics and metabolic flux analysis. The above activities support the ultimate goal of real-time, continuous quality and process verification in pre-engineered, modular and turn-key multi-product manufacturing facilities. For many platforms, such designs can also be imagined in either ready-to-use, microenvironment-based flexible factories or pre-assembled streamlined suite pods promoting closed processing within open-production halls or unclassified “ballroom” or “dance-floor” CNC suites. Such facilities might possess advanced in-line testing technologies, eventually establishing a global, enterprise process control integrating even the scheduling and management of such activities as raw material sourcing and equipment calibration.
- Antibodies: Medical Applications, Immunotherapy and Immune Checkpoints, Anti-Cancer Antibodies, Antibody Humanization Technologies, Biomarkers Development
Chair
William Whitford
GE Health Care, USA
Co-Chair
Yu-Chan Chao
Academia Sinica, Taiwan
Session Introduction
Winfried Stöcker
Euroimmun AG, Germany
Title: Multiparameter autoantibody screening in the diagnosis of neurological autoimmune diseases
Time : 10:55-11:20
Biography:
Winfried Alexander Stöcker is the founder of company EUROIMMUN, CEO. His University studies is from Maximillian University of Wuerzburg, Germany, Doctoral thesis from Institute of Medical Radiology, Maximillian University of Wuerzburg, Germany and Qualification as doctor of laboratory medicine from 1977-1983.
Abstract:
Introduction: The aspect of autoimmunity in neurological disorders attracts a steadily growing interest. Identified autoantibodies include those against intracellular antigens (biomarkers of paraneoplastic neurological syndromes) and newly, those against neuronal cell surface proteins, many of them being pathogenic in several forms of autoimmune encephalitis. However, differential diagnosis of the clinical conditions is challenging due to similar symptomatic appearances and often requires consideration of a broad range of anti-neuronal antibodies. Here, we report on diagnostic benefits of determining a comprehensive antibody profile by multiparametric testing during requested sample analyses. Methods: 16,741 samples were sent to the Clinical Immunological Laboratory (Lübeck) from April 1st 2012 until March 31st 2013. Irrespective of the parameter requested, all samples were tested for multiple parameters using indirect immunofluorescence on BIOCHIP-mosaics (combining various substrates in one reaction field) and immunoblot. Results: 14.1% of the samples were positive for at least one neurological parameter. About half of the detected autoantibodies (52.1%) were of the IgG class. Among these, antibodies against extracellular antigens were twice as frequent as antibodies against intracellular antigens. 53.4% of the samples were positive for the requested parameter whereas 46.6% of the samples were positive for another but requested parameter. Conclusion: Our results reveal that multiparametric screening of samples can increase the hit rate of positive and diagnostically relevant findings by 87% compared to single testing of requested parameters. This may also accelerate the diagnostic process. Both aspects significantly support correct and rapid diagnoses of neurological autoimmune diseases, which is crucial to initiate appropriate and often life-saving therapies.
Jerzy Karczewski
Fraunhofer USA
Title: Plant-based expression of therapeutic monoclonal antibodies targeting viral and bacterial infections
Time : 11:20-11:45
Biography:
Dr. Karczewski received his Ph.D. in Biochemistry from Medical University of Lodz, Poland., After coming to the US he joined Biochemistry Department at Merck Research Laboratories, West Point Pa., where he made numerous contributions to drug discovery efforts, including discovery of novel natural product- and antibody-based inhibitors of intergrins and invented high throughput screening methods which led to discovery of novel inhibitors of cardiac ion channels and other pharmaceutically relevant targets. In 2009 Dr. Karczewski moved to Vaccines Basic Research (also at Merck Research Laboratories) where his research was aimed at discovering novel recombinant vaccines and biologics against bacterial and viral pathogens, including Clostridium Difficile, Chlamydia, RSV, CMV and Neisseria meningiditis. In 2014 Dr. Karczewski joined Fraunhofer Center for Molecular Biotechnology where he supports several projects focusing on plant-based vaccines and monoclonal antibodies.
Abstract:
2014 Liberia Ebola outbreak and subsequent treatment of two aid workers with an experimental cocktail of plant made pharmaceuticals (PMP) brought the attention of plants as a source for therapeutic proteins to the world. Expression of antibodies in plants is not novel. One of the first reported cases was in 1989, but widespread acceptance and recognition of plants as a viable manufacturing system for recombinant protein production has grown greatly in the last decade. Fraunhofer Center for Molecular Biotechnology uses transiently transformed Nicotiana benthiamana plants to express and manufacture recombinant full length murine and human antibodies for use as reagents and therapeutics. The transient transformation expression planform allows for quick screening, scale-up and pilot-scale production in a multi-use facility in rapid succession. Plants harbor no known human pathogens that add a level of safety over traditional expression systems. In addition, plant growth, antibody production and purification are performed without introduction of animal based products. Examples of plant produced mAb against viral and bacterial pathogens will be reviewed.
Lumelle Schneeweis
Bristol-Myers Squibb, USA
Title: Characterization of a PEGylated single-domain antibody
Time : 11:45-12:05
Biography:
Lumelle Schneeweis has completed her PhD from the University of Pennsylvania in Biochemistry & Molecular Biophysics. She has 12 years of pharmaceutical industry experience predominantly focused on protein therapeutics. She has published more than 13 peer-reviewed publications and 2 US Patents. She is currently a Senior Research Investigator at Bristol-Myers Squibb.
Abstract:
Domain antibodies (dAbs) are single immunoglobulin domains that form the smallest functional unit of an antibody. Data on the behavior of these small proteins when covalently attached to the polyethylene glycol (PEG) moiety that is necessary for extending the half-life of a dAb shows the effect of the 40 kDa PEG on hydrodynamic properties, particle behavior, and receptor binding of the dAb. Both ensemble solution and surface methods [light scattering, isothermal titration calorimetry (ITC), surface Plasmon resonance (SPR)] and single-molecule atomic force microscopy (AFM) methods (topography, recognition imaging, and force microscopy) have been used to characterize these conjugates. The large PEG dominates the properties of the dAb–PEG conjugate such as a hydrodynamic radius that corresponds to a globular protein over four times its size and a much reduced association rate. We have used AFM single-molecule studies to determine the mechanism of PEG-dependent reductions in the effectiveness of the dAb observed by SPR kinetic studies. Recognition imaging showed that all of the PEGylated dAb molecules are active, suggesting that some may transiently become inactive if PEG sterically blocks binding. This helps explain the disconnect between the SPR, determined kinetically, and the force microscopy and ITC results that demonstrated that PEG does not change the binding energy.
K E Khalid
Albaha University, Albaha-Saudi Arabia
Title: The presence of interleukin 18 binding protein auto-antibodies and isoforms in chinese patients with rheumatoid arthritis
Time : 12:05-12:30
Biography:
Khalid Eltahir Khalid, PhD, is affiliated as an Associate Professor of Biochemistry at the Department of Basic Medical Sciences, Faculty of Applied Medical Sciences, Albaha University, Saudi Arabia. He hold his BSc in Biochemistry from Omdurman Islamic University, Faculty of Basic Medical Sciences-Sudan, and PhD from Gezira University, School of Medicine and the Institute of Health Sciences, Shanghai Institute of Biological Sciences and Chinese Academy of Science, under supervision of Prof. Jingwu Z. Zhang, and his Post-doctorate in the Molecular Immunology of Leishmania at Sao Paulo University, School of Medicine, Department of Biochemistry and Immunology, under supervision of Prof. Joao Sanatana da Silva. He is a Faculty member at the University of Gezira, performed different activities at the Faculty of Medicine, National Cancer Institute (NCI), and Blue Nile Research Centre for Communicable Diseases (BNRCCD). He attended short courses in Molecular Biology, Immunology, and conducting seminars and teaching activities for under-and Post-graduate Medical students in Sudan. He has been awarded TWAS-CAS fellowship for PhD degree in China, and CNPq-TWAS Post-doctoral fellowship in Brazil. He is currently reviewer for Gezira Journal of Health Sciences, Member of the Brazilian Society of Immunology (Sociedade Brasileira de Imunologia (SBI)), and Trainee Member of Clinical Immunology Society (CIS). He has over 40 scientific papers published and attended several conferences and workshops abroad.
Abstract:
Objective: Human IL-18BP gene encodes at least four distinct isoforms (IL18BPa-d) derived by alternative splicing. Their presence in RA local inflammation is not yet examined. This study aimed to determine the messenger transcript and protein levels of IL-18BP isoforms in patients with Rheumatoid Arthritis (RA). Materials: The study comprises 65 rheumatic patients, 22 Osteoarthritis (OA) and 40 sex and age matched normal controls (NC). Peripheral blood mononuclear cells (PBMCs) and synovial fluids mononuclear cells (SFMCs) were prepared by using Ficoll-Hypaque procedure. The expression and presence of different isoforms were determined by using real-time PCR and ELISA respectively. Results: IL-18BP messenger transcript has been extremely expressed in synovial fluids (SF) and synovial tissues (ST) of RA patients compared to OA patients (p<0.001). IL-18BP autoantibodies were noticed in RA plasma and SF (41.7%; 37.9%), in OA-SF (9.0%), and in plasma of NC (4.0%). Comparable to different isoforms, isoform “c” showed significant local expression (p<0.001) in RA-SFMC and systematic expression (p<0.001) between RA and NC-PBMCs, isoform “a” was least expressed. Isofom “c” and “d” proteins were solely detected by western blot in RA. Conclusions: This study emphasizes the local existence of isoform “c” and “d” and the possible presence of autoantibodies against IL-18BPa in RA patients, which made a pea for further investigation, putting in place their actual role.