8:35 - 9:05 Screening with Crypreserved Cells
Louise Stjernborg, Ph.D., Team Leader, Central Cell Facility, Lead Generation Department, AstraZeneca R&D Mölndal

9:05 - 9:35 An Approach to Post-Phase 1 Cell Line Development
Linda Francullo, Senior Research Scientist 1, Drug Substance Development, Wyeth BioPharma
The biopharma industry has experienced a trend towards a more rapid and constrained, "Platform" approach to Phase I manufacturing process development in order to enable faster clinical development of new product candidates and minimize the investment at risk prior to clinical proof of concept. As a result, it is expected that there will typically be significant need/opportunity for post-Phase I process improvements prior to commercialization, including cell line optimization to achieve increased productivity and/or enhanced product quality. This presentation will describe the current Wyeth BioPharma approach to post-Phase I cell line development, including case studies demonstrating the impact of increased selective pressure, cell adaptation and additional subcloning on cell line stability, productivity and product quality end-points.

9:35 - 10:05 Generation of Stable, High MAb Expressing CHO Cell Lines Using the ACE System
Malcolm Kennard, Ph.D., Director of Cell Line Engineering, Chromos Molecular Systems Inc.
This talk will include a description of the ACE (Artificial Chromosome Expression) system and its application to the generation of recombinant protein expressing mammalian cells. Dr. Kennard will discuss the unique features, timing and advantages of the system and contrast ACE with other expression systems. Case studies will be presented in which CHO cell lines express industry relevant levels (0.5-1.2 g/L) of monoclonal antibodies under non-optimized/non-fed batch shake flask conditions without selection. He will also present data on cell lines that are stable for over 20 generations. Dr. Kennard will also discuss data on fed batch bioreactor studies that show increased expression by 2-3 fold.

11:15 - 11:30 SimCell" MicroBioreactor Platform for High Throughput Cell Culture Process Development: Demonstrated Scalability and Reproducibility 
James Hope, PhD, Vice President, Biotechnology, BioProcessors Corporation

11:30 - 12:00 Production of an IgG1-IL12 Fusion Protein
David Jones, Ph.D., Group Leader, Protein Production, Antisoma plc
AS1409 comprises the humanised antibody BC1 (which binds an oncofetal isoform of fibronectin) fused to interleukin 12 (IL12). IL12 is a heterodimeric protein composed of a p40 and a p35 subunit. An NS0 cell line was generated which first expressed the p40 subunit of IL12; this was then transfected with a plasmid which expressed both the BC1 light chain and the BC1 heavy chain fused to the p35 subunit of IL12. The resulting hexameric protein of approximately 270kDa has been manufactured for clinical trials, and productivity in excess of 550mg/L is achieved in fed-batch bioreactors. A case study will be presented describing USP and DSP process development for this large and complex protein.

12:00 - 12:30 Rational Optimisation of Protein Expression
Loïc Glez, Biotech Engineer, Serono Pharmaceutical Research Institute
Identification of the recombinant protein expression rate-limiting steps is key for the rational improvement of secreted protein expression. This requires complete analysis of the protein expression process, from the transfection efficiency to the final protein secretion. Here we proposed an integrated analytical and quantitative strategy in order to unravel the rate limiting steps in transient gene expression. This strategy applied on a principal set composed of 60 secreted proteins has generated a large amount of information about the protein transient expression in HEK293/EBNA cells. These findings orient the recovery strategies in order to optimize the recombinant protein expression.

12:30 Computer Aided Multi-Parameter Gene Design: Impact of Synthetic DNAs on Protein Expression Enhancement
Professor Dr. Ralf Wagner, Chief Executive Officer and CSO, GENEART AG
Cellular protein production is dependent on various factors, including the underlying nucleotide sequence. GENEART’s optimization algorithm is dedicated to improve expression properties of transgenes by codon adaptation to individual hosts through increasing RNA production, stability and nuclear export by RNA secondary structure removal, CG-value adjustment, splice site avoidance and instability element elimination. In higher eukaryotes, the overall CpG content was demonstrated to be crucial for the level of transgene expression. We report the intragenic CpG-dinucleotide dependent expression of differently designed synthetic genes including cytokines, GFP and HIV-1. Expression yields were monitored on protein and RNA level. A clear correlation of CpG-content, cytoplasmic mRNA levels and protein yields was demonstrated. With the maximum of CpGs, expression yields increased by more than 100% compared to simple codon usage adaptation, and several-fold compared to wild type.

12:45 - 2:00 Lunch on Your Own (Sponsorship Available)

2:05 - 2:35 Reducing Lot-to-Lot Variations in an Insect Medium by Decreasing Yeastolate Content Through Basal Medium Optimization
Steven C. Peppers, Ph.D., Principal Scientist, Invitrogen Corporation
Lot-to-lot variations in cell growth within serum-free insect media have been a problem generally attributed to differences in yeastolate (YE) lots. To address this, we systematically enhanced our base medium, decreased the amount of YE and introduced synthetic lipids. The optimization resulted in lower coefficients of variation with Sf9 and Sf21 cells among different YE lots and higher overall growth and production capacities.

2:35 - 3:05 Growth Medium Optimization for PER.C6 Cells
Sandy McNorton, Research Scientist, R&D, SAFC Biosciences
The PER.C6™ cell line is derived from human embryonic retinoblastoma cells. This cell line is currently licensed by and available through Crucell in the Netherlands. It is used as an expression platform for the development and manufacture of monoclonal antibodies. SAFC Biosciences™ has developed a robust growth medium to support the Crucell platform. This presentation will detail the optimization of this medium which is designed to quickly adapt from serum-free or serum containing media in the PER.C6™ parental cells as well as a large variety of clones developed from the parental cells.

3:45 - 4:15 The Why, When, and How of an Automated Cell Culture System Implementation
Jean Philippe Stephan, Ph.D., Scientist, Assay and Automation Technology Department, Genentech, Inc.
Although cell-based assays have moved into a modern era, cells are still grown and maintained like decades ago: MANUALLY. This step is now a significant bottleneck for many organizations where cell-based assays are a critical path. This presentation will review the different steps involved with the implementation of an automated cell culture system

4:15 - 4:45
Controlling Properties of Cells in Culture Using Microfluidically Patterned Substrates
Jan H. Hoh, Ph.D., Department of Physiology, Johns Hopkins University School of Medicine
Eukaryotic cells in culture respond to the composition and spatial organization of proteins on the substrate, and both the structure and the function of cells can be modulated by controlling the local distribution of extracellular matrix proteins. A number of different approaches to patterning proteins have been developed, with different strengths and weaknesses. We will discuss a new microfluidic approach that allows for highly flexible direct "writing" for compositionally and spatially complex patterns with applications in cell culture.

4:45 5:15 Automated Perfusion System for Homeostatic Cell Culture
Dr. H Joon Paek, Senior Scientist, Regenerative Cell Based Therapies, Tissue Genesis Inc.
We developed an automated cell culture system that allows perfusion, closed-circuit feeding, and temperature and pH monitoring. Compared to the traditional static culture method, perfusion and continuous replenishment of the medium facilitate homeostatic culture and approximates physiological conditions.

5:15 - 5:45 Cell Culture Scale Up and Design
Wei Huang, Director of Process Engineering, Fluor Corporation
As the biotech industry matures, companies are evolving from research and development focused business into more clinical and commercial manufacturing driven business models. The challenge of scale-up cell culture processes from bench/pilot scale to production scale requires good understanding of the cell culture process itself as well as knowledge of equipment design and operation logistics. Latest technology breakthroughs in cell culture development have put more pressure on consistency and robustness of large-scale cell culture in order to meet the high cell density and high titer requirements. The speaker will talk about the critical process parameters unique to cell culture during scale up. Some of the equipment and facility design challenges will be also be discussed in detail.