Cambridge Healthtech Institute’s 9th Annual
Optimizing Cell Line Development
Enhancing Expression
Part of CHI's Ninth Annual The Bioprocessing Summit

August 24-25, 2017 | Westin Copley Place | Boston, MA


The Optimizing Cell Line Development conference brings together cell line development experts who share real-world strategies for quickly achieving cell lines that meet bottom-line goals. Their practical and detailed insights illustrate how to best optimize codons, construct vectors, and how to select and engineer clones and host systems, while maintaining stability and consistency. Gaining a greater understanding of cells through sequencing and the omics sciences will also be addressed, including the CHO genome research, glycoengineering, systems biology, assays, and pathway delineation. In addition, challenges for introducing new technologies will be discussed, along with an overview of industrial trends and regulatory perspectives.

Final Agenda

Thursday, August 24

11:30 am Registration Open

12:15 pm Enjoy Lunch on Your Own

1:15 Dessert Refreshment Break in the Exhibit Hall and Last Chance for Poster Viewing

CELL LINE DEVELOPMENT FOR THE BIOPHARMA INDUSTRY

1:55 Chairperson’s Remarks

Michael Butler, Ph.D., Distinguished Professor Emeritus, University of Manitoba, and CSO, Cell Technology, National Institute of Bioprocessing Research & Training (NIBRT)

 

2:00 KEYNOTE PRESENTATION:

Efficient Cell Line Development for Production and Regulatory Success

Pamela Hawley-Nelson, Ph.D., Associate Director, R&D; Head, Cell Biology in Process Cell Culture, MedImmune, Inc.

2:45 Implementation of New Technologies in Cell Line Generation -- New, Unpublished Data

Jan Schouten, Ph.D., Principal Scientist at Synthon Biopharmaceuticals BV
High-producing, clonal cell lines are at the basis of a biopharmaceutical production process. Strategies will be discussed to obtain cell lines with a high probability of monoclonality in a short time-frame. In addition, state-of-the-art Next Generation Sequencing tools will be discussed to detect and de-select sequence variants in an early stage.

3:15 Transient Protein Production: Harmonizing the Process from Construct Generation through Protein Characterization

Richard Altman, MS, Scientist V, Protein Technologies, Amgen

A robust, flexible transient protein production facility provides critical support to drug discovery efforts. We will review the ongoing evolution of our protein production endeavors focusing on two critical components. The first is the strategic assembly of mammalian expression “tools” that gives us a toolbox capable of expressing diverse and challenging candidate proteins. The second is the harmonization of the entire protein production process thereby reducing turnaround times and increasing throughput.

3:45 Replacing Resistance Genes with Non-Coding RNA for Cell Line Selection

Ted Eveleth, CEO, HocusLocus Inc

We will present a new and novel approach to cell line selection being developed in conjunction with the Austrian Center of Industrial Biotechnology. This approach replaces the traditionally used resistance genes with an siRNA coupled to the gene of interest transcription and performs selection using transfected mRNA.

4:00 Refreshment Break

DEVELOPING CHO CELL LINES

4:15 Sequencing the CHO DXB11 Genome Reveals Regional Variations in Genomic Stability and Haploidy

Christian Schrøder Kaas, Ph.D., Postdoctoral Researcher, Expression Technologies, Novo Nordisk A/S

The copy number from each gene in the genome was calculated from next-generation sequencing data revealed an unexpected degree of haploidy in Chinese Hamster Ovary (CHO) cells. The data can further be mined in order to reveal areas of the genome shown to be stable such as chromosome one and four, which can be hypothesized to host favourable landing platforms for targeted integration of transgenes encoding coagulation factors or antibodies.

4:45 Genome Editing, NGS, microRNAs and Beyond: Transforming New Technologies into More Efficient CHO-K1 Manufacturing Cell Lines -- New, Unpublished Data

Simon Fischer, Ph.D., Scientist, Boehringer Ingelheim Pharma GmbH & Co KG

Genome editing, next-generation sequencing (NGS) and cell line engineering using microRNAs (miRNAs) have emerged as key technologies towards successful biopharmaceutical process development. We have evaluated and integrated these tools into our state-of-the-art cell line development and characterization processes in order to enhance manufacturing of innovative protein therapeutics. This talk will summarize our recent achievements in using these technologies and thus present novel solutions for future challenges in bioprocessing.

5:15 Close of Day

6:009:00 Recommended
Dinner Short Course*

SC 9: Transient Protein Production in Mammalian Cells

* Separate registration required

Friday, August 25

8:00 am Registration Open and Morning Coffee

GLYCOSYLATION

8:25 Chairperson’s Remarks

Richard Altman, MS, Scientist V, Protein Technologies, Amgen

8:30 Cell-Based Glycoengineering of Biologics

Claus Kristensen, Ph.D., Associate Professor, Cellular and Molecular Medicine, Copenhagen Center for Glycomics, University of Copenhagen

Glycan structures are important for efficacy of glycoprotein therapeutics. Glycodisplay have developed a cell based platform to display different glycan structures on drug candidates for optimization. We apply targeted cell engineering to generate large panels of glycoengineered cell lines with different glycosylation capacities. By expressing a drug candidate protein in a panel of our cell lines, we can produce glycovariants of the protein for screening and identification of novel glyco-optimized lead. Applications of our GlycoDisplay technology will be presented.

9:00 Strategies for the Control of Glycosylation during the Production of a Recombinant Protein

Michael Butler, Ph.D., Distinguished Professor Emeritus, University of Manitoba, and CSO, Cell Technology, National Institute of Bioprocessing Research & Training (NIBRT)

Glycosylation is an important post-translational process that leads to heterogeneity of recombinant proteins. This heterogeneity affects the biological activity and clinical efficacy of these bioproducts. The challenge in a bioprocess is to control the heterogeneity to ensure consistency and a profile that maximizes the desired biological properties. The profile can be controlled by cellular glycoengineering, media manipulation or enzymic re-modelling during the downstream process. Possible strategies for the use of each of these methods will be shown for the production of an antibody.

9:30 Optimization and Control of Human Glycosylation during Biotherapeutic Development

Rainer Stahn, Ph.D., Director, Process Development, Glycotope GmbH

A glycoengineered human production platform for the production of human biopharmaceuticals was generated. Based on this technology high yield production of, e.g., mAbs, bispecifics, blood factors and hormones with optimized human glycosylation was established. Aspects of controlling and optimizing glycan structures will be presented at different stages of development as cell line development or process development. Additionally, we present state-of-the-art tools for glycan analysis. Additionally, state of the art tools for glycan analysis will be presented, which is a foundation for rapid and efficient cell line and process development.

10:00 Networking Coffee Break

GLYCOSYLATION & CLONE SELECTION AND OPTIMIZATION

10:30 Investigating Alterations in the CHO N-Glycome Following Exposure to Leachables from Single-Use Bioreactors -- New, Unpublished Data

Jonathan Bones, Ph.D., Principal Investigator, Characterization and Comparability Laboratory, National Institute for Bioprocessing Research and Training (NIBRT)

The cellular N-glycome of CHO cells, characterized using two dimensional liquid chromatography with high resolution mass spectrometry, is highly complex, consisting of a dominant high mannose series and a broad range of complex N-glycans bearing varying degrees of sialylation and poly-lactosamine motifs. The dynamics of the CHO N-glycome is described following exposure to leachable compounds from single use bioreactors. Correlation with product quality attributes, including mAb glycosylation, were also investigated.

11:00 From Thousands of Clones to the Final One

Rolf Köhler, Ph.D., Group Head CLD, Integrated Biologics Profiling, Novartis Pharma AG

CHO cells are the most widely used host for large-scale production of recombinant therapeutic proteins. In order to identify the most suitable lead cell lines, Novartis started to introduce a development process that comprises thorough early candidate profiling focusing on biophysical properties of candidates, followed by a cell line development process applying an efficient selection strategy in combination with process-tailored automation for clone imaging, clone handling, and selection.

11:30 FACS-Based Clone Selection and Host Cell Engineering to Improve the Production and Quality of Recombinant Proteins in CHO Cells

Elizabeth Scheideman, Ph.D., Director, Cell Line Development, Vaccine Production Program, Vaccine Research Center, National Institutes of Health (NIH)

We have been working towards improving our clone selection process and assurance of clonality through the development of a FACS-based platform for single cell cloning, which involves optimization of sorting conditions, medium development, and work on novel reporter-based systems for sorting mAb and especially non-mAb targets. In addition, we are working on methods of modifying our host cells to improve product quality attributes and working to incorporate these metrics earlier in our clone selection process.

12:00 pm Sponsored Presentation (Opportunity Available) 

12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Session Break

INNOVATING CELL LINE DEVELOPMENT

1:25 Chairperson’s Remarks

Jonathan Bones, Ph.D., Principal Investigator, Characterization and Comparability Laboratory, National Institute for Bioprocessing Research and Training (NIBRT)

1:30 BESTcell: Fast Cell Line Development for CHO Clones with High-Yield Protein Production Using Euchromatin-Containing BAC Expression Vectors

Anton Bauer, Ph.D., COO, The Antibody Lab GmbH

Upon stable cell line generation, chromosomal integration site of the vector DNA has a major impact on transgene expression. By using chromosomal loci in BACs and random integration into host cell chromosomes, we developed stable high-yield production cell lines at an unprecedented speed. We performed several case studies for CHO production clones, and we established for antibodies and even difficult-to-express proteins generation of production clones within 3 weeks from transfection.

2:00 Optimizing CHO Cell Culture Process through Mechanistic Understanding of Cysteine Capping Modifications for Improving Site-Specific ADCs -- A Case Study & New, Unpublished Data

Xiaotian Zhong, Ph.D., Senior Principal Scientist/Lab Head, Pfizer BioTherapeutic Research

Cysteine (Cys)-based site-specific ADCs allow for a rapid and simple chemical conjugation reaction to attach diverse linkers/ payloads to provide homogeneous ADC products. This presentation will describe a novel discovery on Cys residue’s post-translational modification in stable CHO cells. The so-called Cys-capping modification utilizes a mechanism that is different from the conventional oxidoreductase-mediated process for protein disulfide formation. The unexpected discovery has led to a novel and highly efficient CHO cell culture process for…

2:30 A Novel Preclinical in vivo Mouse Model for Malignant Brain Tumor Growth and Invasion -- A Case Study & New, Unpublished Data

Thomas N. Seyfried, Ph.D., Professor, Biology, Boston College

Glioblastoma multiforme (GBM) is a rapidly progressive disease of morbidity and mortality and is the most common form of brain cancer in adults. Lack of appropriate in vivo models has been a roadblock to developing effective therapies for GBM. A new highly invasive in vivo GBM model is described that was derived from a spontaneous brain tumor (VM-M3) in the VM/Dk inbred mouse strain. The VM-M3 cells are labeled with the firefly luciferase gene allowing detection through bioluminescent imaging.

3:00 Mito-ID Based Mitochondrial Membrane Potential Staining: A Screening Strategy for Cell Line Development Clone Selection -- New, Unpublished Data

Lina Chakrabarti, Ph.D., Scientist, Cell Culture and Fermentation Sciences, Medimmune, Inc.

Using clonal cell lines and pools from cell line development projects, we performed a series of experiments to evaluate mitochondrial membrane potential (MMP) with Mito-ID dye and found that the intensity of MMP fluorescent signal correlated with the titer values from the producer clones. We also determined that cell sorting based on high MMP staining could potentially combine the two steps of cloning and enriching for high producer cells.

3:30 End of Conference

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