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Optimizing Cell Culture Technology - Day 1


Optimizing Cell Culture Technology Banner

Day 1  |  Day 2 |  Speaker Bios 

This meeting explores today’s evolving strategies and technologies for improving the robustness of mammalian cell cultivation. From media enhancement and cell growth to scaling up, automation, and manufacturing, this leading cell culture meeting is designed to provide helpful information to optimize the job of growing cells.

Optimizing Cell Culture Technology includes talks from industry experts that address critical issues in an atmosphere that encourages sharing, learning, and networking. The agenda again features the very popular small-group breakout discussions that provide the opportunity to discuss important topics with your peers in a fun and collegial setting. In addition, case studies will be presented that shed light on improving yield while maintaining product quality within the regulatory framework.


Scientific Advisors

Susan Dana Jones, Ph.D., Vice President and Senior Consultant, BioProcess Technology Consultants

Dennis Kraichely, Ph.D., Associate Director, CMC Team Lead, Portfolio Management & Technical Integration, Johnson & Johnson Pharmaceutical R&D, Inc.

Robin Ng, Ph.D., Senior Bioengineer, Cell Culture Process Development, Shire Human Genetic Therapies

Fubao Wang, Ph.D., Senior Research Fellow, Vaccine & Biologics Research, Merck & Co.


Monday, August 23 

12:00pm Conference Registration


1:30 Chairperson’s Remarks

Susan Windham-Bannister, Ph.D., President & CEO, Massachusetts Life Sciences Center  Biography


1:40 Opening Keynote Presentation:

John MottStrategies for Evolving a CHO-Based Expression Platform

John E. Mott, Ph.D., Director, Bioprocess R&D, Cell Line Development & Global Biologics, Pfizer, Inc.

There are a multitude of diverse challenges facing scientists working in the mammalian-based biotherapeutic protein expression field today.   Expectations for shortening development timelines, decreasing resources, increasing volumetric titers and reducing the cost of clinical manufacturing continue to intensify.  This presentation will review the various strategies used by Pfizer’s Cell Line Development group in order to meet these expectations.  The presentation will describe a CHO-based monoclonal antibody platform that uses ACF media, automation, cell line engineering and process technologies that enable the rapid identification of cell lines to be used in clinical and commercial manufacturing.  Biography

2:10 Improving Biotherapeutics Through the Use of RNAi

David Kocisko, Ph.D., Principal Scientist, Alnylam Pharmaceuticals

RNAi technology has the ability to improve biotherapeutic manufacturing by selectively and potently turning off problematic genes in biologic producing cell lines. siRNAs target a specific mRNA sequence in the gene of interest to promote its degradation and reduction of corresponding protein synthesis. One of the many strengths of this approach is that siRNAs can be used with any existing cell line, including previously banked cell lines, as an additive to the culture media without any need for cell engineering. In addition, multiple genes can be simultaneously knocked down using a mixture of individual siRNAs directed against each gene target. Using DG44 CHO cells as a model system, siRNAs added directly to the media reduced Bax, Bak, and LDH protein levels by >90% and nearly doubled the integral cell area. Efficient uptake of siRNAs in 40L bioreactors containing DG44 CHO cell cultures has been demonstrated. Finally, siRNA approaches to manipulate other important pathways are currently being developed and demonstrated in DG44 cells.  Biography

2:40 Development of a Glutamine Synthetase (GS) Knockout CHO Host Using Zinc Finger Nucleases

Lianchun Fan, Ph.D., Research Scientist, BioProcess Research & Development, Eli Lilly and Company

Over the past decade, the GS-CHO expression technology has been applied successfully for the production of therapeutic proteins, and in particular, monoclonal antibodies. Selection of recombinant cell lines is based on expression of the gene coding for the glutamine synthetase (GS) enzyme, which is introduced by the expression plasmid. However, parental host cells possess an endogenous copy of the glutamine synthetase gene, which can impact the selection process. In order to eliminate the endogenous GS basal expression and attempt to tighten selection of the transfected population of cells, we utilized zinc finger nucleases targeted against the GS gene to disrupt the genomic loci. The resulting cell lines were characterized to confirm this gene disruption and subsequently evaluated for their performance in the cell line generation process. Based on these studies, the GS knockout cell lines resulted in the elimination of non-producing cells and a significant shift in the mean productivity of the transfected cell population, thus providing for a much more efficient cell line generation screening process.  

3:10 Poster Highlight

3:30 Networking Refreshment Break with Exhibit and Poster Viewing

4:15 Moderated Small-Group Breakout Discussions

Join with your colleagues to discuss the crucial issues associated with culturing mammalian cells and purifying proteins using affinity tags. Small-group discussions are a great way to network, exchange information, and develop collaborations. Each table topic is facilitated by an expert in the field, and discussions can be lively and spirited.

1.   Making a Healthier Cell Line

Moderator:  Lianchun Fan, Ph.D., Research Scientist, BioProcess Research & Development, Eli Lilly and Company

This discussion will mainly focus on the host cell engineering work that deals with anti-apoptosis CHO cell line development.
   • Current strategies to develop anti-apoptosis cells
   • Best targets  to work with
   • Feasibility for new cell lines to be implemented into manufacture process

2.    Alternative Expression Systems

Moderator:  John H. Chon, Ph.D., Senior Director, BioProcess R&D, PERCIVIA, LLC

Chinese hamster ovary (CHO) cells are undisputedly the most widely used of all cell lines in industry for the expression of recombinant proteins for human therapeutics today.  Being a very conservative industry, the biopharmaceutical world has thus far been reluctant to explore and utilize other cell lines despite some obvious advantages over CHO cells.  However, as we are now well into the 21st century, more and more alternative cell lines that address some of the shortcomings of CHO cells are now gaining wider acceptance.  This breakout session will discuss these alternative expression systems and what they bring to the table with respect to safety, quality, and productivity.
   • What are the pros and cons of using CHO cells for human therapeutics manufacturing?  Are any of the cons show stoppers?
   • What alternative cell lines are available to address the “cons” of CHO cells?
   • What are the (perceived and proven) risks of using non-CHO cell lines?  What would be needed to overcome these risks?
   • What is the “dream cell line”?

3.    From Bench to Bedside: Cell Culture's Journey from the Lab to the Clinic

Moderator:  Sajjad Ahmad, Ph.D., Academic Clinical Lecturer, Institute of Human Genetics and North East England Stem Cell Institute, Newcastle University

Cell therapy, for example involving stem cells, is becoming increasingly discussed and indeed used as a means for curing diseases. Bone marrow transplantation is the classic historic example. The aims of this workshop are to discuss how cells processed or cultured in the laboratory can be taken forward to the clinic for therapeutic applications. The following areas will be discussed:
   • Good manufacturing practice
   • Regulatory challenges
   • Scaling up of cells
   • Transportation of cells

4.    Improved Early Screening of Cell Lines

Moderator:  Brian Majors, Ph.D., Scientist, Clinical Cellular Engineering, Biogen Idec

Cell line development groups have made great strides in production cell line titers.  A challenge now is to find methods of early prediction of cell line production and product quality characteristics. This small-group discussion will examine high-throughput screening technologies in cell line development, including:
   • automation,
   • deep well culture, and
   • high-throughput analysis.

5.    Cell Bank Stability:  What Does It Mean and How Do You Measure It?

Moderator:  Jim Moldenhauer, M.S., Senior Research Scientist, Central Cell Banking, Manufacturing Science & Technology, Eli Lilly & Company

The ICH Topic Q 5D, “Quality of Biotechnological Products: Derivation and Characterisation of Cell Substrates Used for Production of Biotechnological/Biological Products”, provides guidance on demonstrating stability of cell substrates both during production and storage.   This guidance is found in section 2.3.3, “Cell Substrate Stability”, and is concerned with evaluation of cell line (or strain) stability during cultivation for production as well as providing evidence for stability of cell banks during storage.   As cell banks have come under increasing regulatory scrutiny, this guidance is referenced during every regulatory agency inspection, and is part of every regulatory submission for product marketing.   Therefore, compliance with this guidance is paramount to commercial manufacturing of biological products.   However, there is no roadmap to success and approaches will vary on how to meet the spirit of these guidelines.    This break-out session will initiate a dialogue between participants on how they believe ‘cell substrate stability’ should or could best be demonstrated in the context of ICH guidance.

6.    Cell Culture Supplements: Quality Aspects

Moderator: Fouad Atouf, Ph.D., Scientific Liaison, The United States Pharmacopeial Convention

One of the challenges associated with the development of therapeutic products derived from cell culture is the use of reagents and ancillary materials as cell culture supplements. This type of reagents can be of animal origin, are not intended to be in the final products and their removal from the finished product is critical for the quality of this category of therapeutics. This small group discussion will focus on:
   • Qualification of cell culture supplements 
   • Animal-derived vs. animal-free materials
   • Residual testing

7.    Dealing with Cell Death in Mammalian Cell Cultures

Moderator: Christopher D. Gregory, Ph.D., CSO, ImmunoSolv Limited; Professor, Inflammatory Cell Biology; Deputy Director, MRC Centre for Inflammation Research, Queen’s Medical Research Institute, University of Edinburgh

Cells die for a variety of reasons in culture, from accidental to deliberate (following drug selection, for example). Dying and dead cells affect their culture microenvironment in different ways and cell death can be a significant limiting factor in effective production, storage and delivery of cells for multiple purposes including bioprocessing, R&D and therapy. This breakout session will discuss issues surrounding death of cells in vitro and measures to minimise its detrimental effects.
   • Causes of cell death in culture
   • Assessing cell death
   • Phases of cell death and impact on the cell culture microenvironment
   • Dead cell removal from cultures
   • Cell death during cell storage and transport

5:15 Roundtable Report Out – Each topic leader will present a brief report of the group’s discussion to the meeting delegates.

5:30 Reception in the Exhibit Hall (Sponsorship Available)

6:30 End of Day One

Day 1  |  Day 2 |  Speaker Bios 


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