Cambridge Healthtech Institute's Fourth Annual

CHO Cell Lines

( CHO细胞株 )

Enhancing Expression, Performance & Process

2018年1月10日 - 11日

 

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

Enhancing Expression through Engineering

2:00 Chairperson's Opening Remarks

Bjorn 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_LewisNathan 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_KristensenClaus 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_StocklLars Stockl, 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

Improving Product Quality through Process

8:15 Chairperson's Remarks

Gyun Min Lee, Ph.D., Professor, Biological Sciences, KAIST

KEYNOTE PRESENTATION

8:20 Advances in Protein Purification

John Kawooya, Ph.D., Director, Biologics Optimization, Amgen

9:00 Speed to IND: Alignment and Acceleration of Critical Early Phase Activities

Kyle_ZingaroKyle 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 High-Throughput Cell Line Screening Specific for Perfusion Processes

Jean-Marc Bielser, MSc, Research Scientist, Biopharma Technology and Innovation, EMD Serono

EMD Serono is following the global trend of continuous manufacturing. To support perfusion cell culture development, small-scale semi-continuous screening methods were developed. These methods were applied on a set of different clones and compared to real continuous conditions (lab-scale bioreactors). Growth and productivity performance were compared and helped to define a ranking strategy for screening application, specific to perfusion.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

11:00 Glycosylation Flux Analysis of Immunoglobulin G Production Using Chinese Hamster Ovary Cell Culture

Rudiyanto_GunawanRudiyanto Gunawan, Ph.D., Assistant Professor, Institute for Chemical and Bioengineering, ETH Zurich

In this work, we developed Glycosylation Flux Analysis (GFA) for analyzing the flux distribution in glycosylation network of monoclonal antibodies. The application of GFA to Immunoglobulin G (IgG) production using Chinese Hamster Ovary cell cultivations revealed (global) cell metabolic and (local) enzyme-specific alterations in the glycosylation network. The GFA pointed to galactosylation as the key regulator and thus a potential target for controlling IgG glycoform distribution in this system.

11:15 Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Therapeutic Protein-Producing CHO Cells

Gyun_Min_LeeGyun 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 (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing

Protein Production: Transient, Stable or Both?

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_PanavasTadas Panavas, Ph.D., Associate Director, Discovery Research, Alexion Pharmaceuticals

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_AltmanRichard 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 Sponsored Presentation (Opportunity Available)

3:35 Refreshment Break in the Exhibit Hall with Poster Viewing

4:15 Stable CHO Pool Expression on Steroids (Using Transposon-Mediated Gene Integration)

Gavin_BarnardGavin Barnard, Ph.D., Group Leader, Biotechnology Discovery Research, Eli Lilly and Company

We describe the development and application of a transposon-mediated gene integration system (TMI) to create stable CHO pools. We demonstrate the superiority of TMI relative to random gene integration (RI), the current method of choice used by the biopharmaceutical industry. CHO pools yielding mAb titers as high as 7.6 g/L were generated using TMI. This represents a 3- to 10-fold increase relative to RI for a panel of molecules.

4:45 The Stability of CHO Genome: Essential for Cell Line Characterization or Not?

Noriko_YamanoNoriko 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.

5:15 Automated Protein Production with the ExpiCHO Expression System

Joseph_DuqueJoseph Duque, Senior Scientist, Merck Research Laboratories

Combining the ExpiCHO protein expression system with automation enables a high throughput protein production platform that can be utilized to generate a variety of proteins in a short amount of time. Here we discuss the automated systems implemented to minimize the manual steps involved in the protein production process as well as the process development findings in the efforts to try to maximize protein expression from the ExpiCHO system.

5:45 Close of CHO Cell Lines Conference

* 活动内容有可能不事先告知作更动及调整。