CHO cells’ rapid rise in production prominence is due to their adaptability to various culture conditions, gene plasticity, and ability in proper folding, posttranslational modifications, and glycosylation of desired proteins. Thus, advances in
CHO cell lines and culture continue to significantly improve biotherapeutic production. This achievement is due to progress in engineering stable and transient cell lines, enhancing cell culture conditions and performance, as well as optimizing process
development. When all are accomplished, higher-production titers and better product quality result. The CHO Cell Lines conference gathers cell line engineers, cell culture specialists and bioprocess development managers to explore
the latest data, tools and strategies for improving protein expression, production, and product quality.
Final Agenda
WEDNESDAY, JANUARY 10
1:00 pm Registration
2:00 Chairperson’s Opening Remarks
Bjørn Voldborg, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
2:05 Optimizing Expression of Proteins in CHO through a Systems Biology Approach
Nathan E. Lewis, Ph.D., Assistant Professor, Department of Pediatrics, University of
California, San Diego
In our lab, we are mapping out the cell pathways controlling CHO cell growth, protein synthesis, and protein glycosylation. Here I discuss our work in which we have developed computational models to predict the cell costs for protein synthesis and identify
how to improve protein synthesis through media and genetic modifications.
2:35 Optimizing Biologics by Cell-Based Glycan Display
Claus Kristensen, Ph.D., CEO, GlycoDisplay Aps
Glycan structures are important for efficacy and distribution of biologics. Optimization of glycans has been hampered by inefficient technologies for glyco-engineering in mammalian cells. Now GlycoDisplay offers technologies allowing development of novel
glyco-optimized biologics. GlycoDisplay has applied targeted cell engineering to generate cell lines with different glycosylation capacities. By expressing a drug candidate protein in panels of glycoengineered cell lines, followed by screening novel
glyco-optimized leads are identified.
3:05 Refreshment Break in the Exhibit Hall with Poster Viewing
4:00 Overexpression of Ebola Virus Envelope GP1 Protein
Zhongcheng Zou, Ph.D., Staff Scientist, Structural Immunology Section, Lab of Immunogenetics, NIAID/NIH
To elucidate the role of the mucin-like domain of GP1 in Ebola-host attachment and infection and to facilitate vaccine development, we constructed a GP1 expression vector containing the entire attachment region. Cysteine 53 of GP1 was mutated to serine
to avoid potential disulfide bond mispairing. Stable expression clones using codon optimized open reading frame were developed in human 293-H cells with yields reaching ~ 25 mg of GP1 protein per liter of spent medium.
4:30 Using GlycoExpress to Overcome Production Limitations for Difficult-to-Express Proteins
Lars Stöckl, Ph.D., Director, Glycoprotein Development and PTM Analytics, Glycotope GmbH
Even though productivity for CHO systems has remarkedly improved over the last years, some biopharmaceuticals like bispecific constructs or complex glycoproteins remain very challenging. We present case study data from clone and upstream perfusion development
for the human GlycoExpress cell line, which overcomes productivity limitations.
5:00 Expression of Recombinant Blood Coagulation Factor VIII: Importance for Human Healthcare and Approaches to Improve the Protein’s Yield and Quality
Andrey G. Sarafanov, Ph.D., Chemist, Principal Investigator, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration (FDA)
Deficiency in factor VIII (FVIII) results in abnormal bleeding (Hemophilia A), which is treated by infusions of FVIII. However, the FVIII production is challenging as the protein is expressed at low levels both in plasma (0.3 nM) and heterologous
systems. The presentation overviews approaches to improve FVIII production, in particular, re-design of the protein and its gene, optimization of the protein expression and purification, and selection of test methods.
5:30 - 6:45 Networking Reception in the Exhibit Hall with Poster Viewing
6:45 Close of Day
THURSDAY, JANUARY 11
7:45 am Morning Coffee
8:15 Chairperson’s Remarks
Gyun Min Lee, Ph.D., Professor, Biological Sciences, KAIST
8:20 Integrating Cell Culture with Magnetic Protein A Bead-Trap Technology Accelerates Antibody Purification
John K. Kawooya, Ph.D., Director, Biologics Optimization, Discovery Research, Amgen
Antibody engineering produces large numbers of molecules (200-500 molecules at 30-50ml each) which require purification, analysis and screening for potency, binding, pharmacodynamics, pharmacokinetics, manufacturability, expression levels and
stability in order to select leads. Ever since its inception over 30 years ago, the AKTA system combined with Protein A agarose columns has remained the “workhorse” of antibody purification from cell cultures. However, the inability
of this system to process multiple samples in parallel coupled with both its limiting flow rates, its requirement for multiple FTEs to remove cells and particulate from each sample prior to loading together with the potential for sample swapping
errors and cross contaminations – all impose major bottlenecks in expediting large purified panels of molecules. In this presentation, I show how a single FTE with parallel Magnetic Protein A bead-trap technology accelerates delivery
of high-quality purified antibodies in high yield directly from small (30ml-5liters) to large (25-liter wave-bag) crude cell cultures without centrifugation or filtration.
9:00 Speed to IND: Alignment and Acceleration of Critical Early Phase Activities
Kyle Zingaro, Ph.D., Development Scientist II, Early Stage Development, Alexion
Pharmaceuticals
Speed to IND is the current battle cry across early phase biologics development. Although this faster-is-better approach comes with some risk, new technologies and alignment of workflows can afford both faster and better decisions during this
crucial phase of new product development. This is especially true across the Discovery and Process Development handoff including the interface of discovery, preclinical, process, formulation, and analytical teams. Here we present new data
and approaches to improve that handoff and detail the impact on timelines and quality of molecules and cell lines in early phase development.
9:30 "Lost in Translation": Bridging the Gap between Academia and Biotech
Tsafi Danieli, Ph.D., Director, BioGiv Excubator & Head, Protein
Expression Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem
One of the most difficult and frustrating aspects of basing a startup company on academic findings is translating and transferring academic findings to biotech language. This procedure is often frustrating to both parties and requires psychological
skills as well as critical review of the research. Working in the interphase between academia and industry, there are several preemptive strikes we can take to avoid some of the major pitfalls.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
11:00 Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Therapeutic Protein-Producing CHO Cells
Gyun Min Lee, Ph.D., Professor, Biological Sciences, KAIST
Host cell proteins (HCPs) accumulate extracellularly during the cultures of recombinant CHO (rCHO) cells, potentially impairing product quality. HCPs accumulated extracellularly in batch and fed-batch cultures of rCHO cell lines were identified
and quantified by mass spectrometry. This dataset of HCPs provides insights into determining the appropriate target proteins to be removed during both the cultures and purification steps for ensuring good therapeutic protein quality.
11:30 Using SUREscan™ to Survey Genetic Changes in Stable CHO Cell Lines
Pierre-Alain Girod, CSO, Selexis
CHO cells are the most frequently applied host-cell system for industrial protein therapeutic manufacturing. Rapid generation of high-producing clones that don’t lose expression capability over time has been a major industry focus.
Using SUREscan™ with next-generation sequencing (NGS), we can quickly analyze whole genomes of any cell line, improving traceability of Research Cell Banks (RCBs). In contrast to other CHO published data, we will show that SUREtechnology
Platform™ generates RCBs with chromosomally stable lineages.
12:00 pm Session Break
12:15 Luncheon Presentation: Get Your High Protein Concentrations Right on the Money with Lunatic
Thomas Martens, Principal Scientist, Unchained Labs
Stop quantifying proteins one by one hoping that your old reader is getting the numbers right. Get rid of that dilution step you always need to measure 200 mg/ml IgG or even higher. Come learn about how Lunatic gets rid of all dilutions,
eliminates any risk of cross contamination and accurately measures protein concentration at high throughput and high concentrations. We’ll talk about how lunatic: • measures either 16 or 96 samples in one run • uses
only 2 µL for each measurement • needs only 5 minutes • Requires no dilutions
1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Remarks
Saurabh Sen, Ph.D., Principal Scientist, Biotherapeutics Discovery, Boehringer Ingelheim
2:05 A High-Density CHO-S Transient Transfection System: Comparison of ExpiCHO and Expi293
Tadas Panavas, Ph.D., Senior Principal Scientist, Biotherapeutics Molecule
Discovery, Boehringer Ingelheim Pharmaceuticals, Inc.
Chinese Hamster Ovary (CHO) cells are the principal mammalian host used for stable cell line generation and biotherapeutic protein production. Until recently, production of milligrams to grams of protein in CHO transient systems was challenging.
To overcome such challenges, we evaluated the ExpiCHO system, a high-density CHO-S transient transfection system, and compared it to the Expi293 and FreeStyle MAX CHO transient systems. Detailed analysis was performed on protein titer,
monodispersity, enzyme activity, and posttranslational modifications.
2:35 Transient Protein Production: Harmonizing the Process from Construct Generation through Protein Characterization
Richard Altman, MS, Scientist, Protein Technologies, Amgen
A robust, flexible transient protein production facility provides critical support to drug discovery efforts. We 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.
3:05 Difficult to Express Proteins: Novel Plasmid Technology to Significantly Increase Product Yield in CHO Cells
Marco Cacciuttolo, Ph.D., Head of Operations, Batavia Biosciences
Yield is still an area that requires significant improvement for many promising recombinant proteins and antibodies. Novel vector technology enables rapid generation of stable, CHO cell lines able to provide at least 10-fold more product
per cell.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Lost in Translation: On the Formation of Protein Sequence Variants
Zhongqi Zhang, Ph.D., Scientific Director, Attribute Sciences, Process
Development, Amgen
With modern mass spectrometry and appropriate informatics tools, a large number of low-level sequence variants in therapeutic proteins are detected and quantified. This large collection of information allows for a deeper understanding
of the mechanism for the formation of sequence variants, thereby facilitating optimization of cell line and cell culture process to minimize them.
4:45 The Stability of CHO Genome: Essential for Cell Line Characterization or Not?
Noriko Yamano, Ph.D., Senior Scientist, Manufacturing Technology
Association of Biologics; Guest Academic Staff, Graduate School of Engineering, Osaka University
The chromosomes in CHO cells frequently cause genomic variations, due to genetic instability. Distribution and stability of chromosomes were examined in CHO-DG44 cells, and two cell lines expressing different numbers of chromosomes
were isolated from the original CHO-DG44 cell line to investigate the effect of aneuploid cells on recombinant protein production. In addition, gene expression profiles between cells with disparate chromosome numbers have been
compared by mRNA-seq analysis.
View the Interview
5:15 High-Throughput Screening of Transfection Efficiency of dTtaPS Reagent Library, and Its Application for Transient Production of Proteins in Micro Bioreactors
Harsh Jain, Ph.D., Visiting Associate, FDA
5:45 Close of CHO Cell Lines Conference