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

11:30 Main Conference Registration

CULTURING CHO CELLS

1:00 pm Chairperson’s Opening Remarks

Tiffany Rau, Ph.D., Global Technical and Technology Manager, Eli Lilly and Company

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

ANALYSIS APPROACHES & AMBR

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.

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

OPTIMIZING PROCESS DEVELOPMENT

7:55 Chairperson’s Remarks

Martin Heitmann, Ph.D., Senior Scientist, Cell Culture Technology, Novo Nordisk A/S

8:30 Prediction of Cell Culture Performance and Molecule Quality Attributes from Micro-Scale Fed-Batch Cultures

David 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.

9:30 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

METABOLIC FLUX ANALYSIS & QUALITY

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

PERFUSION PRODUCTION

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

Martin 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

Christopher 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

TOOLS AND TECHNOLOGIES TO ENHANCE CELL CULTURE PROCESSES

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 results.

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