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Cambridge Healthtech Institute’s 6th Annual

Optimizing Expression Platforms
( 优化表达平台 )

Tools for Effective Expression, Production, and Purification

2019年1月17日~18日

 

The utilization of engineered therapeutic proteins for basic research, clinical diagnostics, and therapy continues to expand. Consequently, protein expression laboratory managers and researchers face challenges for efficient expression, production, and purification even while improving quantity and quality and minimizing time and cost. Transient protein production (TPP) has the advantage of speed and limiting risk while stable transfection – the longer and more complex process – has the advantage of producing long-term expression of the biotherapeutic of interest. The rapidly increasing need for recombinant proteins necessitates further improvements in both technologies.

Cambridge Healthtech Institute’s 6th Annual Optimizing Expression Platforms conference convenes protein expression specialists who share their experiences with process and platform differences, tradeoffs, and improvements for producing recombinant proteins in their expression and production laboratories.

Final Agenda

THURSDAY, JANUARY 17

7:45 am Registration and Morning Coffee

Improving Production with Stable Cell Lines

8:10 Organizer’s Welcome Remarks

Mary Ann Brown, Executive Director, Conferences, Cambridge Healthtech Institute

8:15 Chairperson’s Opening Remarks

Howard R.G. Clarke, PhD, Principal Scientist, Cell Sciences, Seattle Genetics


KEYNOTE PRESENTATION

8:20 Chromosome Stability Approach: Lengthening of High-Yield Production Levels of IgG-Producing CHO Cells by Downregulation of Breast Cancer 1

Takeshi Omasa, PhD, Professor, Department of Material and Life Science, Graduate School of Engineering, Osaka University

The effects of breast cancer 1 (BRCA1) downregulation on gene amplification efficiency and long-term productivity were investigated in CHO cells. Our results suggest that high-producing cells, which maintain their productivity long term, were efficiently established by BRCA1 downregulation. In this presentation, I would like to introduce the chromosome stability and effect of BRCA1 downregulation.

9:00 Antibody Expression Stability in CHO Clonally Derived Cell Lines and Their Subclones

Howard R.G. Clarke, PhD, Principal Scientist, Cell Sciences, Seattle Genetics

Cell line development involves lengthy screening to identify a stable line having consistent growth, productivity and product quality. To investigate production stability in CHO cells we analyzed primary clones and their respective subclones. Cell lines derived from single cell progenitors grow into populations of cells with phenotypic heterogeneity. Here I present the genetic and epigenetic characterization of these heterogeneous cell line populations.

9:30 Presentation to be Announced

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

11:00 Manufacturing and Characterization of Novel HIV-1 Vaccine Candidates – Success and Challenges

Antu K. Dey, MSc, PhD, Senior Director, R&D, Vaccine Product Development Center, International AIDS Vaccine Initiative (IAVI)

Discovery endeavors in recent years have identified a number of HIV-1 vaccine candidates that showed promising results in preclinical animal studies. These results have prompted process and analytical development efforts to support cGMP manufacturing of some of these HIV-1 recombinant protein vaccine candidates for evaluation in (proof-of-concept) Phase I clinical studies. To express these recombinant vaccine candidates, stable CHO cell line expression was used. Stable CHO cell line development and clonal selection required a ‘functional’ titer assay for selection of top stable clones. The trimeric complex conformation and extensive posttranslation modifications of these vaccine candidates demanded that extensive characterization of the vaccine antigen produced from top clone be performed to ensure appropriate glycosylation, disulfide linkage and high level of percent trimer. Combination of upstream and downstream process development and subsequent extensive characterization led to production of these vaccine candidates with desired attributes and subsequent ‘release’ of drug product for Phase I clinical evaluation.

11:30 Engineering a Stable CHO Cell Line for the Expression of a MERS-Coronavirus Vaccine Antigen

Mun Peak Nyon, PhD, Research Associate, Pediatrics, Pediatric Tropical Medicine, Baylor College of Medicine

Human vaccine against MERS-CoV is not available. We have developed a stably transfected adherent CHO cell line for the production of the MERS-CoV protein subunit, S377-588 (Fc tagged). The adjuvanted protein vaccine expressed in adherent CHO could protect transgenic animal model from infection with live MERS-CoV. We also have developed a suspension monoclonal CHO cell line able to express S377-588-Fc in serum-free media, which is ready for scaled-up production.

12:00 pm Session Break

12:10 Luncheon Presentation I: New Tools for Screening & Harvesting Solutions for CHO & HEK293 Cells, for Both Transient and Stable Cells

Samuel Ellis, Vice President, Thomson Instrument Company

Evaluation of different transfection tools, product quality, and titer for both CHO and HEK293 cell lines. Data will be presented on techniques and technology that mimic large-scale bioreactors in non-controlled devices from 1mL-3L. Technologies presented include well plates and culture tube systems with incorporated filtration methodology. A new direct harvesting technique will also be introduced that eliminates centrifugation while maintaining 0.2um sterile filtration. All of these tools will be presented with case studies from scientists.

Batavia12:40 Luncheon Presentation II to be Announced

 

1:10 Ice Cream Break in the Exhibit Hall with Poster Viewing

Tools for Transient Protein Production (TPP)

2:15 Chairperson’s Remarks

Masamichi Kamihira, PhD, Professor, Faculty of Engineering, Department of Chemical Engineering, Kyushu University

2:20 Scaling Up a High-Titer HEK293 Transient Transfection Process

Tia Arena, MSc, Engineer I, Department of Cell Culture, Genentech

HEK293 transient expression systems are often used to quickly generate protein for research and preclinical studies. Here we describe engineering a HEK293 cell line that is more resistant to apoptosis and shear stress. After process optimization for seed train (35 L) and transient transfections (up to 25 L), this robust cell line-enabled expression of antibodies and non-antibody proteins up to 800 mg/L in 7 days.

2:50 Recombinant Production of the Toxic Anti-Cancer Lectin Viscumin in Tobacco Plants and Microbial Cells: A Comparative Analysis of Yield, Process Costs and Toxicity

Johannes Buyel, Dr. rer. nat., Dr.-Ing., MSc, Head, Integrated Production Platforms, Fraunhofer Institute for Molecular Biology and Applied Ecology IME

Viscumin is a potential anti-cancer protein that cannot be produced in mammalian cells due to its inherent toxicity. Manufacturing in microbial systems is cumbersome due to the formation of inclusion bodies that require a complex process, which provides only low recoveries. Instead, plants can be used as a cost-effective alternative expression system that simplifies production and yields a more active product.

3:20 Sponsored Presentation (Opportunity Available)

3:35 Networking Refreshment Break

4:00 Manipulating Glycan Profile in a Transient Expression System and Application of a High-Throughput Capillary Western Method Using Lectins for Detection

Silvino Sousa, MSc, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center, AbbVie

I discuss approaches for modulating N-linked glycosylation of recombinant therapeutic proteins by manipulating media, process and/or genetics of the host cell factory. What is also needed is a rapid, simple, yet protein- and titer-agnostic method for deriving detailed glycan signature directly and simultaneously from multiple samples of cell culture conditioned medium. I review methods that we have implemented for rapidly screening for glycan signatures directly from cell culture supernatants.

4:30 Accumulative Transgene Integration into a Predetermined Chromosomal Site of CHO Cells

Masamichi Kamihira, PhD, Professor, Faculty of Engineering, Department of Chemical Engineering, Kyushu University

An accumulative site-specific gene integration system (AGIS) based on the Cre-recombinase/loxP system, using mutated loxP sites (Kameyama et al., 2010, Biotechnol. Bioeng., 105, 1106–1114) has been applied for the generation of recombinant CHO cells for producing antibodies (Wang et al., 2016, J. Biosci. Bioeng., 124, 583–590). AGIS can provide an efficient tool for repeated integration of transgenes into a predetermined chromosomal locus.

5:00 PANEL DISCUSSION: Transient, Stable or Both?

Speed, limiting risk and protein quality are often cited as advantages of transient protein production (TPP), while stable transfection – the longer and more complex process – has the advantage of producing long-term expression of the biotherapeutic of interest. The rapidly increasing need for recombinant proteins necessitates further improvements in both technologies.

Moderator:

Masamichi Kamihira, PhD, Professor, Faculty of Engineering, Department of Chemical Engineering, Kyushu University

 

Panelists:

Tia Arena, MSc, Engineer I, Department of Cell Culture, Genentech

Johannes Buyel, Dr. rer. nat., Dr.-Ing., MSc, Head, Integrated Production Platforms, Fraunhofer Institute for Molecular Biology and Applied Ecology IME

Howard R.G. Clarke, PhD, Principal Scientist, Cell Sciences, Seattle Genetics

Silvino Sousa, MSc, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center, AbbVie

5:30 Close of Day

FRIDAY, JANUARY 18

8:00 am Registration

8:00 BuzZ Sessions with Continental Breakfast

Protein therapeutics is a fast-growing global market. As the science improves, so does the complexity of the R&D organization. Ensuring product quality plus speed to market requires insights from stakeholders working across the stages of protein science R&D. Join experts representing this PepTalk pipeline, peers, and colleagues for an interactive roundtable discussion. Topics include highlights from the week’s presentations, new technologies and strategies, challenges, and future trends.


Moderator: Richard Altman, MS, Scientist, Protein Technologies, Amgen

 

Managing a Protein Production Lab:
How to Make the Most of Your Resources

9:00 Chairperson’s Remarks

Richard Altman, MS, Scientist, Protein Technologies, Amgen

9:05 Managing a Collaborative Multidisciplinary Laboratory: Challenges, Strategies, and Benefits

Challise Sullivan, Life Scientist III, Advanced Solutions Group, Leidos

The advantages of a laboratory staffed with skilled, versatile personnel and equipped with systems applicable to numerous applications can be vast. Integrating scientific and engineering expertise, cutting-edge technology, and a collaborative team enables innovations spanning a wide range of disciplines and applications beyond those typically attained by conventional academic or industrial practices. This talk encompasses approaches to effectively manage multidisciplinary laboratory teams along with the challenges and benefits of doing so.

9:35 High-Throughput Cloning for Biomarker Discovery and Functional Genomics

Vel Murugan, PhD, MBA, Research Scientist, Virginia G. Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University

We generate and distribute expression clones around the world. DNASU is a central repository for plasmid clones and collections (DNASU.org). Currently we store and distribute over 300,000 plasmids including 75,000 human and mouse plasmids, full genome collections, the protein expression plasmids from the Protein Structure Initiative as the PSI: Biology Material Repository (PSI : Biology-MR), and both small and large collections from individual researchers. We are also a founding member and distributor of the ORFeome Collaboration plasmid collection. We discuss HT cloning methods that we employ for generating expression clones.

10:05 Recombinant Protein Production: Harmonizing the Process from Construct Generation through Protein Characterization

Richard Altman, MS, Scientist, Protein Technologies, Amgen

A robust, flexible 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.

10:35 Networking Coffee Break

11:00 Making More Proteins: How to Get the Work Done and How to Avoid It

Peter Schmidt, PhD, Director, Recombinant Technologies Research, CSL Behring

The increasing number of projects in early R&D combined with the need to characterize and evaluate new approaches and ideas result in a continuously increasing number of requests to purify and QC proteins. In order to meet these demands, it is necessary to find a good balance between available resources and goals that can be realistically achieved.

11:30 CLOSING PANEL DISCUSSION: Protein Production Lab Challenges: Methodologies, Strategies, and the Art of Managing Multiple Projects

There are many challenges in operating protein production labs. This panel focuses on the following topics: initiating projects, basic expression and purification systems, pros and cons for each system, screening platforms, troubleshooting and how much time should be spent on each system before moving to the next option. On top of “hands on” tips, we touch upon strategies on how to manage multiple “top priority” projects.

Moderator:

Richard Altman, MS, Scientist, Protein Technologies, Amgen

 

Panelists:

Vel Murugan, PhD, MBA, Research Scientist, Virginia G. Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University

Peter Schmidt, PhD, Director, Recombinant Technologies Research, CSL Behring

Challise Sullivan, Life Scientist III, Advanced Solutions Group, Leidos

Silvino Sousa, MSc, Senior Scientist, Global Protein Sciences, AbbVie Bioresearch Center, AbbVie

Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

12:30 pm Close of Conference

* 活动内容有可能不事先告知作更动及调整。

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