Cambridge Healthtech Institute’s 11th Annual
Optimizing Cell Culture Technology
Enhancing Knowledge for Growing Cells
August 3-4, 2015
Part of CHI's 7th Annual The Bioprocessing Summit
August 3-7, 2015 | Westin Copley Place Hotel | Boston, Massachusetts
This leading cell culture meeting explores today’s evolving strategies and technologies for improving mammalian cell cultivation, including omics technologies and Synthetic Biology. Industry experts offer insights into optimizing conditions, as
well as, cell biology in an effort to improve yield, while also addressing where cell culture is heading as it meets ever greater production demands and quality standards.
The agenda again features a session focused on CHO (Chinese Hamster Ovary cells), as well as small-group breakout discussions that provide the opportunity to discuss important topics with your peers from around the world.
Monday, August 3
8:00 am Pre-Conference Registration and Morning Coffee
9:00-11:30 Recommended Short Course
Optimizing Media - Achieving Super Soup
11:30 Main Conference Registration
1:00 pm Chairperson’s Opening Remarks
Tiffany Rau, Ph.D., Global Technical and Technology Manager, Eli Lilly and Company
1:10 OPENING KEYNOTE PRESENTATION:
Getting to the Root of the Problem: What are the Fundamental Factors Limiting Growth and Productivity in CHO Cell Cultures?
Gregory Hiller, Ph.D., Associate Research Fellow, Culture Process Development, BioProcess R&D, Pfizer, Inc.
Why do they stop growing?!? We will describe experimental strategies undertaken over the last eight years in my group to answer this most basic question and understand the underlying cellular metabolism. We will also explain our methods of overcoming
the problem of growth cessation of CHO cells which have enabled dramatic increases in overall culture productivity in the industry standard fed-batch culture and in less common modes of bioreactor operation.
1:45 Development and Manufacturability Assessment of Chemically-Defined Medium for the Production of Protein Therapeutics in CHO Cells
Wai Lam W. Ling, Ph.D., Senior Principal Scientist, Process Development & Engineering, Biologics BioProcess
Development, Merck Research Labs
Internally developed chemically-defined (CD) media offers flexibility for protein production process development. Through DOE screenings and component optimization, a CD basal medium (CDM) was developed for CHO cell culture. Culture performance of CDM
manufactured by vendors was poor compared to in-house preparation. An investigation revealed key medium components were sensitive to commercial manufacture. CDM was subsequently reformulated into a manufacture-compatible core medium with sensitive
components supplemented separately.
2:15 Host Cell Protein Expression during Extended CHO Cell Culture
Kristin Valente, Ph.D., Associate Principal Scientist, Merck
As the biopharmaceutical industry moves towards continuous bioprocessing, it is important to consider the impact of extended CHO culture on extracellular HCP expression, particularly for impurities that are difficult to remove during purification. Proteomic
techniques were applied to evaluate HCP expression over 500 days of culture. The effect of cell age on the HCP impurity profile is presented and implications of variable HCP expression on downstream purification are discussed.
2:45 Refreshment Break
3:15 Optimizing Cell Culture Processes and Associated Analytical Methods
Lada Laenen, Ph.D., Senior Director, Manufacturing Science and Technologies, Genzyme
3:45 Enhanced Process Consistency, Robustness and Success in Cell Line Selection Using High-Throughput Bioreactors
Wenqi Xie, Associate Scientist III, Technical Development, Biogen, Inc.
AMBR is now a common high-throughput bioreactor option for process development and cell line selections. At Biogen, we evaluated the robustness of our platform process in AMBR and determined important levers that affect cell culture performance and product
quality attributes. Moreover, through systematic retrospective and prospective evaluations of cell line screening results from multiple projects, we proposed the optimal number of cell lines to screen and therefore recommend using high-throughput
bioreactors to achieve target titer in cell line selections.
4:15 Breakout Discussions
This session provides the opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish collaborations
or commiserate about persistent challenges. Then continue the discussion as you head into the lively exhibit hall for information about the latest technologies.
Biosimilar Development: Cell Line Screening and Process Optimization – The Right Balance?
Moderator: Arnaud Périlleux, Assistant Project Manager, Biotech Process Science, Merck Serono SA
Approaches to Establishing Product Specifications from Early Clinical Development to Commercial
Moderator: To be Announced
Integrating Continuous Up- and Downstream Processing to Increase R&D Protein Production
Moderator: Martin Heitmann, Ph.D., Senior Scientist, Cell Culture Technology, Novo Nordisk A/S
Novel Production Systems and Regulatory Considerations
Moderator: Tiffany Rau, Ph.D., Global Technical and Technology Manager, Eli Lilly and Company
Feeding Strategies for High Density CHO to Keep Cells Growing Longer in Log Phase.
Surfactants, how much and what is the best?
Moderator: Sam Ellis, Vice President, Biochemist, Thomson Instrument Co.
- Pluronic vs. Simithecone; does your media have enough surfactant for high density CHO
- Amount of Surfactant needed?
- Transient effects?
- Stable CHO needs for surfactants?
- Glucose, Glutamate, cell boost, what should we do?
Solving Development Issues with Large-scale Transient Transfection
Moderator: James Brady, Ph.D., Vice President, Technical Applications and Customer Service, MaxCyte
- De-risk development with transient transfection of the manufacturing cell line
- Shorten development timelines with large-scale transient transfection
- Scale-up/Scale-down of transient transfection
- Using Stable cell lines versus transients downstream
5:15 Discussion Report-Outs
5:30 Grand Opening Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
Tuesday, August 4
7:30 am Registration and Morning Coffee
7:55 Chairperson’s Remarks
Martin Heitmann, Ph.D., Senior Scientist, Cell Culture Technology, Novo Nordisk A/S
8:00 FEATURED PRESENTATION:
Delivering the Pipeline: A Global Perspective of Process Development and Design for Manufacturing
Tiffany Rau, Ph.D., Global Technical and Technology Manager, Eli Lilly and Company
8:30 Prediction of Cell Culture Performance and Molecule Quality Attributes from Micro-Scale Fed-Batch Cultures
Brühlmann, MSc., Biotech Technology and Innovation, Biotech Process Sciences, Merck Serono SA
The selection of a fed-batch cultivation system is often based on throughput and cost. However, the process knowledge derived from different systems and scales is not necessarily identical. Hence, a careful characterization of systems which are already established or newly implemented is essential. Here, we present the cell culture performance and molecule quality attributes of 12 different recombinant CHO cell lines expressing the same antibody in shaking 96-deepwell plates operated in fed-batch mode. We subsequently cultivated the same recombinant cell lines with the same expansion and fed-batch protocol in systems which are typically used in cell culture process development: 50 mL vented shake tubes, micro- and lab-scale bioreactors. The results of this study show the potential of shaking 96-deepwell plates in predicting cell culture performance and molecule quality attributes at larger scales. The system can be used in high throughput mode at very early stages of process development to screen clonal cell lines or to study the impact of media components.
9:00 Novel Virus-Like Particle and Nanoparticle Vaccine Production – An Overview of Cell Culture Process Development
Payal Biswas, Ph.D., Scientist, Upstream Process Development, Vaccine Production Program, Vaccine Research Center/NIAID/NIH
In order to rapidly produce novel VLP vaccines and nanoparticle influenza vaccines for Phase I clinical trials, a mammalian cell culture-based, robust transient transfection platform process was developed. Transfection conditions and process parameters
were optimized to achieve a viable yield for clinical production and scaled up to 50L fully disposable bioreactor system for GMP manufacturing. Background and supporting data for Phase I manufacturing of these vaccines will be presented.
Manufacturing Core Competency for Biosimilar Contenders
Shun Luo, Ph.D., CEO & President, Jianshun
Biosciences Co., Ltd.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
10:30 Applying MFA to the Process Development Environment
Neil Templeton, Ph.D., Senior Scientist, Bioprocess Development,
Merck Research Laboratories
Metabolic flux analysis (MFA) was applied to multiple fed-batch processes. A stoichiometric analysis was then applied to identify potential nutrient limitations to biomass and antibody synthesis. Finally, multivariate data analysis
(MVDA) was applied to an experimental time-course of metabolic fluxes, in an effort to identify fluxes of a fed-batch culture that correlated best with final titer. All of this work was conducted in a fashion to suit the rapid timelines
of industrial process development.
11:00 Towards Cell Engineering Using 13C-Metabolic Flux Analysis and Insight from Cancer Metabolism
Woo Suk Ahn, Ph.D., Postdoctoral Associate, Chemical Engineering, Massachusetts Institute of Technology
Cancer metabolism is a promising topic to be utilized for cell engineering field. Cancer cells quickly produce bioenergy and anabolic precursors by aerobic glycolysis that is increased significantly under hypoxia. Here, we quantified cancer
metabolism using 13C-metabolic flux analysis and validated by gene knock-down/overexpression techniques. We revealed how rapidly growing cells coordinate metabolism to survive and proliferate under hypoxic condition.
11:30 5-Hydroxymethylcytosine (5hmC) Barcoding and Cell Identity
Richard R. Meehan, Ph.D., Project Leader, Chromosomes and Gene Expresion, MRC Human Genetics
Unit, IGMM, University of Edinburgh
An assumption underlying the use of cultured cells is that they retain and mimic the molecular characteristics of the tissue from which they were derived or conform to an industry standard that enables their reproducible use in bio-manufacturing.
By studying the establishment of fibroblasts and T-cells in culture, we found that adaptation resulted in a rapid and comprehensive re-programming of the transcriptome and epigenome, indicative of an altered cell state. Re-programming
involved almost complete loss of 5hmC in cultured cells. Restoration of 5hmC profiles is indicative of restoration of cell identity.
12:00 pm From Upstream to Downstream: Large-Scale, High Titer Transient Transfection Platform for Biomanufacturing
James Brady, Ph.D., Vice President,
Technical Applications and Customer Service, MaxCyte
Harmonizing upstream and downstream processes can reduce the time it takes to move a biotherapeutic to market. In this presentation, data developed with MaxCyte’s flow electroporation, closed, single-use, cGMP-compliant system will
demonstrate large-scale transient transfection in CHO cells and biologically relevant cells and the resulting high titers. The results demonstrate the platform’s scalability, high transfection efficiency, and cell viability.
The outcome is a cost-efficient, timely high yield and a harmonization between upstream and downstream.
12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:15 Session Break
1:55 Chairperson’s Remarks
Wai Lam W. Ling, Ph.D., Process Development & Engineering, Biologics BioProcess Development, Merck Research Labs
2:00 Scale-Down Tools for Evaluation of Perfusion Cultivations
Heitmann, Ph.D., Senior Scientist, Cell Culture Technology, Novo Nordisk A/S
Strategies are presented to enable screening of multiple process parameters at different scales, while being focused on simple, yet predictive, scale down models for perfusion processes. Due to the difficulty of implementing cell retention
in small scale, a chemostat based scale-down system was developed. The conception of a repeated exponential fed-batch scheme to mimic chemostat cultivations is described and its implementation in 10 mL scale ambrTM system cultures
is presented. This system is then compared to classical bench top chemostat and ATF perfusion cultures.
2:30 Progress towards Efficient Implementation of Continuous Upstream Processes in Early Development
Daryl Powers, Ph.D., Senior Scientist,
Early Cell Culture Development, Sanofi Global Biotherapeutics
We have implemented several strategies to streamline early process development for perfusion production of a therapeutic IgG that is being readied for transfer to the pilot plant for clinical material generation. These strategies include
some perfusion-specific techniques in addition to many of the same tools and methods that are used for fed-batch process development.
3:00 Optimizing Osmolality and Cell Expansion Strategy to Improve Cell Culture Performance in a Perfusion-Based Bioreactor Process
Rives, Ph.D., Senior Upstream Development Engineer, BioProcess Development, Shire plc
During development of a perfusion-based bioreactor process for production of a therapeutic protein, a significant amount of variability in bioreactor terminal viable cell density and viability profiles was observed. This presentation describes
the approaches taken to investigate the potential root causes of performance variability and summarizes the key experimental findings. In addition, the proposed control strategy will be shared.
3:30 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Biologics Data Platform for Tailored Support of Cell Line Development
Christian Bender, Ph.D., Computational Biologist, Global Drug Discovery, Global Biologics, Bayer HealthCare
We have successfully implemented the Biologics Data Platform (BDP) for tailored support of our screening and protein production processes. In the context of our cell line development process, we present the integration of BDP with our
automation workstation. We demonstrate the power of using a comprehensive data management platform to track data for cell line clones and fed-batch experiments together with molecule information such as primary sequences and experimental
4:45 Portable X-Ray Fluorescence Spectrometer: A Tool for Biopharmaceutical Forensic Investigations
Jessica Mondia, Ph.D., Research Scientist, Biogen Idec, Inc.
A portable X-ray fluorescence (XRF) spectrometer is a small, cheap, easy and fast instrument for multi-elemental analysis. In biopharma, forensic investigations usually refer to determining the root-cause and evaluating the risks associated
with deviations from GMP guidelines including batch records, procedures and SOPs. Here we introduce the use of a XRF spectrometer for biopharmaceutical forensic applications as an in-house-portable diagnostic tool to help resolve or
guide investigations in a timely fashion.
5:15 Close of Conference