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Overview | Short
Courses | Day 1 (Joint Session) |
Day 2 | Day 3 | Download Brochure
7:30am – 4:00pm Registration Open
7:30 Breakfast Workshop (Sponsorship Available)
Production Strategies
8:15 Chairperson’s Remarks
8:20 High-Throughput Cloning and Over-Expression of Integral Membrane Proteins
Philip Gao, Ph.D., Director, Protein Production Core Laboratory,
University of Kansas
Production of membrane proteins is a critical challenge. We describe a fast, reliable, and economical protocol for preparing high quality, homogeneous samples of helical integral membrane proteins. Over-expressed proteins in the membrane or in the form of inclusion bodies were solubilized using minimal amount of denaturant in the presence of detergent. Solubilized proteins were purified using one step immobilized metal affinity chromatography. This streamlined sample preparation protocol has been combined with screens for detergents and sample conditions using NMR spectroscopy to produce high quality proteins.
8:50 Strategic Approach for Balancing Speed-to-Clinic and Commercialization Objectives During Antibody Development
Kevin M. Bailey, Ph.D., Vice President, Preclinical Manufacturing and Process Development, Regeneron Pharmaceuticals, Inc.
Development of antibody manufacturing processes requires a fine balance between the need-for-speed to gain early access into the clinic versus the need to have an optimized cell line and manufacturing process suitable for commercialization. Regeneron has developed a flexible approach to meet both of these needs without delay of clinical development timelines. Considerations in the introduction of a new cell line and process modifications during late-stage clinical development will be discussed along with the positive impact on commercial manufacturing capacity and cost-of-goods.
9:20 Frequent FDA Inspection Observations - Protein Manufacturing
Mark A. Lynch, Vice President, PAREXEL Consulting
Presented by a former senior FDA enforcement official, this session recaps the most frequent citations listed by FDA resulting from adverse inspection observations. The nature of the citation, common causes, and steps to prevent receiving adverse observations will be discussed. The presentation also covers best practices for responding to adverse inspection observations through corrective action plans.
9:50 Coffee Break in the Exhibit Hall
10:45 Poster Awards
Optimizing Production
11:00 Transforming a Standard Multiwell Plate into a Bioreactor – Recombinant Protein Production at High Cell Densities with EnbaseTM
Peter Neubauer, Ph.D., Professor, Bioprocess Engineering, Technical University of Berlin
EnbaseTM or enzyme based substrate delivery is a fed-batch derived technology for shaken culture systems, e.g. 96-well and deep well plates. This very robust technology allows a controlled growth of cells to high cell densities, just like in a bioreactor and the growth rate is simply controlled by the amount of an initially added biocatalyst. Yields of cells and products have at least a 10 fold increase over traditional techniques. EnbaseTM is an enabling technology for high-throughput approaches, such as screening of enzyme or phage display libraries, small scale production of proteins for crystallization and labelization purposes, as will be demonstrated in the talk. As EnbaseTM is a fed-batch process, process optimization is simply possible with multiwell plates with a possibility for direct scale-up into the bioreactor, which significantly reduces time and cost of process development.
11:30 Antibody Fragment Therapeutics - Approaches to Development of an Integrated Production System
John Liddell, Ph.D., Head, Process Science, Research and Development,
Avecia Biologics
The success as Mabs as therapeutics has been the main area of growth in the area of therapeutic proteins for almost 10 years now. This success arises from their high levels of specificity for the relevant disease targets with this high level of specificity has allowed them to meet unmet medical need as well as reducing harm to healthy cells typically resulting in fewer adverse effects compared to other medicines. Antibody fragments being minimal binding fragments derived from whole antibodies are predicted to be the next major class of therapeutic proteins to be
commercialised. As bio-therapeutics they offer the potential to provide new therapies arising from altered specificities and small molecular size. New opportunities for production of antibody fragments compared to Mabs arise as they can be produced by microbial technology. This has the opportunity to reduce manufacturing complexity compared to Mab production by cell culture. The small size of antibody fragments provides challenges in achieving in vivo pharmakinetics and conjucation to PEG (polyethylene glycol) can be applied to reduce turnover. The presentation will describe data from Avecia's development of an integrated approach to all stages of antibody fragment production including new approaches to fermentation, purification and specific
pegylation.
12:00 Establishment of an Industrial Production Process for Recombinant ACE2
Manfred Schuster, Ph.D., Chief Scientific Officer, Apeiron Biologics
Angiotensin converting enzyme 2 (ACE2) is a promising candidate for an enzyme substitution therapy to treat acute cardio-vascular, pulmonary and inflammatory diseases. It is a highly
N-glycosylated Zink-metalloprotease expressed on lung, kidney, heart and liver cells whose expression is
down-regulated in several life-threatening diseases. We have completed at present our pre-clinical development phase and will enter clinical trials in Europe, Canada and USA very soon, both in healthy volunteers as well as in ARDS patients. This talk focus on the outcome of our meetings with regulatory agencies in Europe and USA and presents the strategy we have adopted to produce, to formulate and to characterize our protein therapeutic. A second part of this presentation summarizes the pharmacological properties of ACE2 and the results of our toxicology and safety pharmacology program.
12:30 Close of Morning Session
12:45 Luncheon Workshop (Sponsorship Available) or
Lunch on your Own
Shaping the Future of Protein Production
2:00 Chairperson’s Remarks
2:05 High-throughput Cell-free Expression to On-demand Protein Microarray Technologies
Deb K. Chatterjee, Ph.D., Associate Director, Protein Expression Laboratory, National Cancer Institute, SAIC-Frederick, Inc.
Availability of thousands of full-length clones following whole genome sequencing of various organisms has led to the development of new methods and novel technologies to study function of the proteins. We have developed an improved cell-free protein expression technology that enable us to produce almost 10 fold more proteins compared to commercially available methods. In addition, we have developed a simple and novel technology where DNA microarrays can be converted to protein microarrays in the chip on-demand. This technology saves time and also, eliminates the cost of protein expression and purification of thousands of mammalian proteins.
2:35 In Vitro Study of Membrane Protein Complexes Solubilized in Nanolipoprotein Particles
(NLPs)
Matthew A. Coleman, Ph.D., Senior Scientist, Biosciences and Biotechnology Division, CMELS, Lawrence Livermore National Laboratory
We have developed cell-free methods for producing soluble membrane protein contained in nanolipoprotein particles (NLPs). NLPs have shown great promise as a biotechnology platform for solubilizing and characterizing membrane proteins. However, current approaches are limited because they require extensive efforts to express, purify and solubilize the membrane protein prior to insertion into NLPs. By the simple addition of a few constituents to cell-free extracts, we can produce soluble membrane proteins in NLPs with considerably less effort. Importantly, the approach also provides a single-step process for the production of functional soluble membrane proteins that eliminates the need for cell growth, cell lysis, and subsequent purification, refolding etc. We have demonstrated this process on multiple membrane proteins important for proton translocation, host-pathogen interactions and cell surface signaling. Our approach represent a unique solution to solubility and purification problems associated with membrane proteins.
3:05 Mapping Ligands and Binding Proteins to Improve Protein Stability and Identify Functional Characteristics
Frank R. Collart, Ph.D., Molecular Biologist, Biosciences Division, Argonne National Laboratory Biosciences Division, Argonne National Laboratory
The ligand binding ability of proteins is coupled to inherent characteristics such as function, cellular localization and biochemical properties. We have assessed the utility of a ligand mapping technique to identify binding ligands for protein families such as transcription factors, sensor proteins, or binding proteins of ABC transporters. Our approach for the identification of bound ligands uses a fluorescence-based thermal shift
(FTS) assay for high-throughput screening of protein-ligand interactions. The FTS approach is a target-independent assay that uses a fluorescent dye to monitor protein unfolding. The assay was validated with a set proteins of known binding specificity and was consistently able to map proteins with their known binding
ligands. The general utility for assessment of binding specificity has been demonstrated for a group of uncharacterized binding proteins for ABC transporters. The assays reveal that these proteins bind a diverse set of ligands representative of the functional and metabolic diversity of this organism. Specific binding was detected for several functional chemical categories including aromatic compounds, polyamines, dicarboxylate metabolites, and amino acids. The specific experimental assignments illustrate the ability of the FTS approach to provide biologically significant and specific annotation and to increase or extend biochemical/biophysical characterization of proteins.
DISPOSABLES
3:35 BioSMB™: A New Continuous disposable chromatography process
Scott Fulton, Founder, Scientific Advisor, Tarpon Biosystems
The downstream processing bottleneck is leading to the exploration of processes with higher efficiencies and higher productivities. BioSMBTM is a technology that refines traditional SMB into a viable option for biopharmaceutical purification. By implementing a fully disposable-format fluid path and modular design, the BioSMBTM technology addresses the key issues associated with biopharmaceutical processing, such as elimination of cleaning validation and rapid campaign change-over. This talk will present experimental data of a MAb downstream process using a bench-scale BioSMBTM system. The presentation covers the capture of MAb from cell supernatant on various commercially available Protein A media, packed in disposable cartridges. The data showed that the BioSMBTM outperformed the batch process in terms of MAb recovery and HCP reduction, while offering a significant reduction in buffer consumption and a three to tenfold increase in productivity. Based on the experimental data, projections will be given for the impact of BioSMBTM technology on large-scale manufacturing of monoclonal antibodies.
4:05 Roundtable Discussion – The Pros & Cons of Disposables
Panelists:
Leigh Pierce, B.S., President, PacificGMP, Inc.
Scott Fulton, Founder, Scientific Advisor, Tarpon Biosystems
Geoffrey Hodge, Ph.D., Vice President, Process Development & Technology, Xcellerex LLC
4:30 Close of Conference
Overview | Short
Courses | Day 1 (Joint Session) |
Day 2 | Day 3 | Download Brochure
For more information, please contact:
Mary Ruberry, Conference Director
Phone: 781-972-5421
E-mail: mruberry@healthtech.com
For exhibit and sponsorship information, please contact:
Suzanne Carroll, Manager, Business
Development
Phone: 781-972-5452
E-mail: scarroll@healthtech.com
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