TUESDAY, SEPTEMBER 18 - DAY TWO

7:30am Morning Coffee (Sponsorship Available)

8:30 am Chairperson's Remarks
Tom Fletcher, R&D Director, Irvine Scientific

8:35 Cell Engineering of Apoptosis
Mohamed Al-Rubeai, Ph.D., Professor, School of Chemical and Bioprocess Engineering, University College Dublin
The identification of apoptosis as the major mechanism of cell death during the production process has raised the importance of studies of cell death and its prevention when implementing culture optimisation strategies. In this talk I will highlight the studies which have demonstrated the induction of apoptosis and methods to identify and analyse cell death. The suppression of apoptosis under these conditions has been demonstrated by a number of recent reports, and I will present ways in which this knowledge may be applied to engineer cells for high resistance to apoptosis.

9:05 An Alternative Method for Cell Line Adaptation
S. Edward Lee, Ph.D., Research Fellow, Pfizer Groton Labs
The standard method for cell line adaptation is by weaning. It is labor intensive and time consuming. We have developed a faster and more direct method to adapt cell lines to growth in serum-free or animal-components-free media. This method also favors the generation of the more stable genotypes as opposed to the less stable phenotypes derived from the weaning method.

9:35 Know Your Cells and What You Want From Them
Linda B. Jacobsen, Ph.D., Senior Manager, Berit Biotech, LLC
Optimizing expression in a misidentified cell line may result in the generation of a wonderful cell line that may not be able to be used. It is critical that cells be properly identified at the start of the research, and procedures be followed to ensure that the cells remain unchanged. Examples of cross-contamination, misidentification and changes with passage level will be presented. Once the cell type is defined, the desired endpoint for the research should be considered during optimization of experimental conditions. Specific examples will be given for transient transfection where media, ratios and amounts of reagent and DNA were optimized at small scale. In these optimization experiments multiple endpoints were used to measure success of transfection since protein production levels do not always correlate with % of cells transfected.

10:05 Using Real Time Metabolic Measurements of Cells to Predict Behavior in a Bioreactor
David Ferrick, Ph.D.,Vice President, Assay Development, Seahorse Bioscience, Inc.
Understanding the effect of culture conditions on mitochondrial respiration and glycolytic flux enables optimization conditions that maximize desirable characteristics, such as increased cell growth and protein production. XF24 Extracellular Flux Analyzer data will be presented showing how this easy to use, label free technology can assist with clone selection and media optimization.

10:20 Networking Coffee Break

10:50 Chairperson's Remarks

10:55 A Systematic and Direct Comparison of Different Translational Optimization Technologies on Protein Expression Using the ACE System
Malcolm Kennard, Ph.D., Director, Cell Line Engineering, Chromos Molecular Systems Inc.
Expression of recombinant genes can be adversely affected by problems at the transcriptional and translational levels. Codon and/or translational optimization attempts to identify these problems within the DNA and generate modified sequences that will minimize these effects and result in increased recombinant protein expression. To test the effects of codon/translational optimization on recombinant protein expression, an IgG4 MAb sequence was optimized using three different algorithms. These optimized sequences were compared by evaluating the MAb productivity of CHO cell lines generated by the Chromos ACE System process. This talk will briefly summarize the ACE System process and detail the comparison of the optimized sequences in ACE System generated cell lines. 

11:25 Routine Maintenance and Banking of Mammalian Cell Lines in Protein- and Peptide-Free Minimal Culture Media
Ferruccio Messi, Ph.D., President & CEO, Cell Culture Technologies
The presentation reports on the selection and authentication of human and animal cell lines that can be routinely maintained and cryopreserved in strictly defined minimal culture environments, i.e. minimal culture media (MCM). The most important feature of these cell lines derived from the serum-dependent parent cell lines Sp2/0, NS0, P3X63.653, COS-1, COS-7, BHK-21, HEK-293, Vero, and CHO-K1 - is their routine maintenance in MCM totally free of proteins, peptones, peptides, chemically undefined additives and animal-derived ingredients. Said MCM exclusively consist of a minimal number of pharmaceutical grade small molecules, all of which are characterized by their respective CAS and EINECS registry numbers. The MCM used for routine maintenance and banking can also be used for producing biologicals such as recombinant proteins, monoclonal antibodies and viral particles with the above cell lines. Except for the CHO-K1 cell line, the MCM adapted cell lines derived from the serum-dependent parent cell lines were authenticated, certified and banked by the European Collection of Animal Cell Cultures (ECACC). While the ECACC currently distributes the newly selected serum-free cell lines worldwide, our company promotes the technology transfer with respect to the development of industrial culture processes with mammalian cell lines in MCM.

11:55 Technology Spotlight 
(Sponsorship Available – Please contact Suzanne Carroll, scarroll@healthtech.com or call 781-972-5452) 

12:10 pm Integrated Approach of Product Characterization for Cell Line Screening & Development
Chrstine P. Chan, Ph.D., Reserach Fellow, BioProcess R&D, Merck & Co., Inc.
In early bioprocess development for therapeutic monoclonal antibodies, various production cell clones and process conditions are evaluated for cell growth characteristics, expression stability, product titers and product quality attributes before final selection of the most desirable clone and process. Typically for the latter application, complementary analytical tools including HPLC, capillary electrophoresis and mass spectrometry are used to monitor product structural heterogeneity for variations in fragment size, charge and oligosaccharide modifications. This presentation will review application examples on characterization of monoclonal antibodies produced from different mammalian cell lines and evaluate the use of key methods in comparison and in combination.

12:40 Lunch on Your Own (Sponsorship Available)

2:00 Chairperson's Remarks

2:05 Manufacturing Commercial Products Derived from Cell Culture Platforms
Sarad Parekh, Ph.D., Director, Cell Culture Process Development, Phyton Biotech Inc.
This presentation will outline Phyton’s cGMP production and operation capability that uniquely positions it as the world leader in plant cell culture technology. To achieve this goal Phyton has brought together the infrastructure and expertise to produce cGMP-grade biopharmaceutical products. This presentation will examine Phyton’s process development and scale-up capabilities that has enabled operating with stirred tank reactors with capacities as large as 75,000 L.

2:35 Fed-Batch vs. Large-Scale Perfusion Culture? Making the Right Choice -- Impact on Scale Up and Manufacturing Cost of Goods
Bo Kara, Director, Science and Technology, Avecia Biologics Ltd.
For a number of therapeutic protein targets, the nature of the product, its complexity, issues with stability, etc ‘drives’ the selection of cell culture perfusion technology as the preferred upstream manufacturing route. During R&D, perfusion systems can offer a simple 'small-scale' system that can provide protein product for preclinical studies. This can often translate into the process used to support early clinical manufacture of cell culture protein products. The rationale for making the decision between fed-batch and perfusion technology will be examined together with an assessment of the impact of product titre on ultimate CoG for the manufacture of a monoclonal antibody product using a dedicated manufacturing facility based on either fed-batch or large-scale perfusion technology.

3:05 Technology Spotlight (Sponsorship Available – Please contact Suzanne Carroll, scarroll@healthtech.com or call 781-972-5452) 

3:20 Networking Refreshment Break

3:45 Novel Approaches for High Throughput Cell Line Development and Selection
Rosemary Drake, Ph.D., Director, Business Development, The Automation Partnership
Selection of the optimum cell line from very large populations is time consuming and laborious. Two new automated cell culture technologies for cell line selection and characterisation enable data to be obtained from much larger numbers of cell lines, allowing users to evaluate a wider range of molecular approaches and cell culture processes in parallel, thereby facilitating more rapid and efficient cell line development. This presentation will show how highly parallel processing can contribute to significantly reduced cycle times and better decision making, through rapid identification of optimum cell lines and culture conditions.

4:15 Bypassing Conventional Approaches for Rapid Discovery of Rare-Event Cell Lines
Christopher J. Mann, Ph.D., Senior Scientist, Biopharmaceutical Technologies, Genetix
The pharmaceutical industry is under pressure to reduce development time and costs for new therapeutic proteins. To address this issue, a single-step technology, ClonePix FL, was developed that screens thousands of cell clones in situ for monoclonal antibody secretion, and quantitatively collects only the best clones based on specificity or rate of production. Case studies will be presented to demonstrate the efficacy of the technology. New applications will also be discussed, which permit the detection and isolation of cells secreting monomeric proteins, and the quantitation of clones based on phosphopeptide specificity. 

4:45 Electronic Cell Sensor Array Technology for Real-Time, Label-Free Live Cell Quality Control
Naichen Yu, Ph.D., Research Field Scientist, Biology, ACEA Biosciences
ACEA Biosciences has introduced the Real-Time Cell Electronic Sensing (RT-CES) system which uses impedance sensor technology to non-invasively quantify cells based on number, morphology, and adhesion quality in real-time. Real-time monitoring of cellular processes by the RT-CES system offers distinct and important advantages in monitoring the quality of live cells for cell-based assays in a label-free environment. The RT-CES system provides high content and information-rich data that is beyond the scope of traditional monitoring or single-point assays.

5:15 pm End of Conference