Optimizing Protein Expression

 

단백질 발현의 최적화

Expression of heterologous proteins presents many challenges and understanding expression systems is key. The 9th annual Optimizing Protein Expression conference delves into protein expression by examining and enhancing expression systems, including CHO and other mammalian systems, E. coli, yeast and baculovirus. What is the best expression system for expressing your protein of choice? Ease and cost of scale-up must be considered to ensure successful bottom-line results. Experts will share case studies and disclose data while divulging details of expression systems’ underlying mechanisms. Comparing and contrasting systems will also be featured to increase understanding in the quest for greater productivity.

Final Agenda

TUESDAY, APRIL 9

Recommended Short Course*

SC9: Introduction to Biophysical Analysis for Antibody Discovery and Development

Christine P. Chan, PhD, Principal Scientist, Global Manufacturing Science & Technology, Sanofi

 

*Separate registration required.

WEDNESDAY, APRIL 10

7:15 am Registration and Morning Coffee

7:25 - 8:25 PANEL DISCUSSION: Women in Science – Inspired Professional and Personal Stories (Continental breakfast provided)

Moderator:

Jennifer-ChadwickJennifer S. Chadwick, PhD, Director of Biologic Development, BioAnalytix, Inc.; Co-chair, Mentors Advisors and Peers Program, Women In Bio, Boston Chapter


Panelists:

Joanna BrewerJoanna Brewer, PhD, VP, Platform Technologies, AdaptImmune


Charlotte A. RussellCharlotte A. Russell, MD, DMSc, Chief Medical Officer, Alligator Bioscience


Susan RichardsSusan Richards, PhD, Presidential Scientific Fellow, Translational Medicine Early Development, Sanofi R&D


Kristi SarnoKristi Sarno, Senior Director Business Development, Pfenex


단백질 생산 전략

8:30 Chairperson’s Opening Remarks

Philippe Billiald, PharmD, PhD, Professor, Biochemistry, University Paris-Sud & Acticor Biotech


8:40 KEYNOTE PRESENTATION: Early Process Development: Challenges and Opportunities for Emerging Therapies

Beaucamp_NicolaNicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development, Roche Diagnostics GmbH

A number of new molecules, different from standard antibody structures, have been advanced into clinics by Roche pRED. In order to discover and develop differentiated biologics, Roche’s strategy is based on engineering technologies leading to new formats and processes which bear several challenges and many opportunities for technical development towards the clinic and beyond.

9:10 Site-Specific Biotinylation of Secreted Proteins in Mammalian Cells in vivo – Made Viable!

Mark Trautwein, PhD, Senior Scientist, Pharmaceuticals R&D, Preclinical Research, Expression Technologies, Bayer AG

Site-specific biotinylation of secretory target proteins is desirable to achieve in vivo during protein expression in mammalian cells. However, conventional techniques like the AviTag suffer from serious losses in expression yields, preventing their applicability as a generic approach. In this presentation, I will describe how we have optimized a protein tag to solve the problem and achieve this goal.

9:40 Expression Optimization of Antibody Fab Fragments in Escherichia coli with Non-Native Amino Acid (NNAA) Incorporated

Harun Rashid, PhD, Senior Principal Scientist, Molecular Technology, Ambrx, Inc.

Ambrx’s site-specific NNAA incorporation technology platform in Escherichia coli uses orthogonal transfer to suppress Amber stop codon for incorporation of a NNAA. In this study, expression of two ‘difficult-to-express’ antibody Fab fragments with NNAAs inserted were systematically optimized by expression vector and strain engineering. Then, the various beneficial components for each Fab were put together into single expression vectors resulting in significant improvements in the expression titer of each Fab over the starting strains.

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

10:15 Women in Science Speed Networking in the Exhibit Hall

CHO 기반 발현의 최적화

10:55 Combining Biophysical Analytics with Next-Generation Sequencing for Deep Characterization of mAb Producing CHO Cell Lines

Thie_HolgerHolger Thie, PhD, Senior Manager, Technology and Innovation, Biologics Development, Boehringer Ingelheim Pharma GmbH & Co. KG

Differences in gene expression can lead to unpredicted behavior of mAb producing CHO cell lines during bioprocess development. We observed remarkable differences in N-linked glycosylation between two production clones derived from the same CHO cell line. NGS-based genetic characterization allowed unraveling the genetic regulation behind the differences in N-linked glycosylation. This study shows how NGS helps to gain a deeper understanding of CHO production cell lines.

11:25 Engineering CHO Cell Lines for the Production of Hard-to-Produce Proteins

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

Using our high-throughput cell line engineering platform, we have engineered CHO cells able to produce therapeutic proteins that have previously not been possible to produce in CHO cells. This approach may result in improved therapeutic proteins with better biological properties, such as increased half-life, improved activity, etc.

11:55 Importance of Appropriately Glycosylated Species, and Modulation Efforts, to Achieve Desired Post-Translational Modifications in CHO-Derived Monoclonal Antibodies

Chauhan_GauravGaurav Chauhan, MS, Associate Principal Scientist, BioProcess Development, Merck & Co., Inc.

The glycosylation profile of the monoclonal antibodies has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during protein production. Since glycosylation is an important parameter for IgG function, several strategies to modify glycosylation profiles of human IgG or to select the most efficient glycoforms have been explored. Further, considerable efforts have been made to understand how these glycoforms can be modulated as desired. I would like to present on current understandings of such efforts.

12:25 pm 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.

12:55 Luncheon Presentation I to be Announced



1:25 Luncheon Presentation II to be Announced

1:55 Session Break

유전자 변형 CHO 세포

2:10 Chairperson’s Remarks

Nicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development, Roche Diagnostics GmbH

2:15 Generation of Superior Host Cell Lines for Biomanufacturing

Laux_HolgerHolger Laux, PhD, Fellow, Cell Line Development, Novartis Institutes for BioMedical Research

CHO cells are the most widely used host for large-scale production of recombinant therapeutic proteins. Using transcriptomic approaches, we have identified target genes involved in productivity and product quality. Subsequently, a variety of novel parental CHO cell lines were generated applying cell line engineering techniques as ZFN or TALEN. These novel knockout CHO cell lines are superior in respect to productivity and/or product quality.

2:45 Achieving Predictable Recombinant Gene Expression in CHO Cells Using CRISPR-Mediated Genome Engineering

Pristovsek_NusaNuša Pristovšek, PhD, Postdoctoral Researcher, The Novo Nordisk Foundation Center for Biosustainability (CFB), Technical University of Denmark

Chinese hamster ovary (CHO) cells are one of the major hosts for production of complex therapeutic proteins. Efficient synthetic biology tools are of great interest to improve production in CHO cell factories. Here, our latest development of these tools will be demonstrated. Together with high-throughput technologies and systems biology approaches, synthetic biology can pave the way toward accelerated generation of desirable CHO cell factories with predicted culture performance.

3:15 Scaling Up and Scaling Out: Pushing the Boundaries of Transient Protein Production

Wilkinson_Ian_MABIan Wilkinson, PhD, CSO, Absolute Antibody, Ltd.

Whilst transient yields have improved drastically in the last decade, scalable systems are time-consuming and costly to implement. Absolute Antibody has developed systems which scale up and scale out protein expression and purification, enabling the rapid and cost-effective production of milligram to gram quantities of large panels of proteins.

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing

4:45 Problem-Solving Breakout Discussions - Click here for details

5:45 Networking Reception in the Exhibit Hall with Poster Viewing

7:00 End of Day

THURSDAY, APRIL 11

8:00 am Registration and Morning Coffee

발현계의 비교/대조

8:30 Chairperson’s Remarks

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

8:35 Microbial or Mammalian Cells for Expression of Therapeutic Antibody Fragments: Making the Choice with the End in Mind

Billiald_PhilippePhilippe Billiald, PharmD, PhD, Professor, Biochemistry, University Paris-Sud & Acticor Biotech

Because antibody fragments are usually aglycosylated proteins, many host cell expression platforms can be used for production, including bacteria, yeast and mammalian cells. However, all these expression systems are not equivalent in terms of cell line development, culture time, product quality and cost of production. We will report differences that may have to be considered before pharmaceutical development and moving forward to the clinic.

9:05 Using Non-Coding RNA for Improving Recombinant Protein Expression and Growth from Mammalian Cells and Microorganisms

Shiloach_JosephJoseph Shiloach, PhD, Director, Biotechnology Core Lab, NIDDK, NIH

Non-coding RNAs, including microRNA and siRNA in eukaryotes, and small RNA in prokaryotes, are regulatory molecules that can affect protein expression through interactions with specific sections of the host mRNA in mammalian cells, and mRNA and proteins in bacteria. The utilization of mammalian microRNA and siRNA to enhance growth and protein expression from HEK and CHO cells and affecting E. coli metabolism using bacterial small RNA will be presented.

9:35 Organize, Optimize, and Measure Biologics R&D with Modern Software

Wickramasekara_Sajith_MABSajith Wickramasekara, CEO and Co-Founder, Benchling

Benchling is a biologics-native informatics platform used by over 160,000 scientists to configure biologics workflows and run day-to-day R&D. This presentation will highlight how Benchling has helped leading biopharma companies to organize, optimize, and measure their biologics R&D.

9:50 Overcoming Limitations of Conventional Tag Systems – Strep-Tactin®XT Applications

Karhaus_Darius_MABDennis Karthaus, MSc, IBA Lifesciences

The Strep-Tactin®XT:Twin-Strep-tag®-purification system enables protein purification at high yields and purity under physiological conditions. Providing the highest binding affinity among all affinity tag systems, the technology fulfills the demands of mammalian expression systems (e.g. Expi) and is well suited for downstream applications like SPR.

10:05 Coffee Break in the Exhibit Hall with Poster Viewing

비포유류 발현계

11:05 Rational Engineering of an Improved Secretion Signal for Pichia pastoris

Glick_BenjaminBenjamin Glick, PhD, Professor, Molecular Genetics and Cell Biology, University of Chicago

The yeast Pichia pastoris is widely employed to secrete heterologous proteins. Typically, the N-terminal portion of pre-pro-alpha-factor is used as a secretion signal. This secretion signal promotes post-translational translocation into the ER, so proteins that fold in the cytosol are poorly secreted. The alpha-factor pro region can also promote aggregation in the ER. We applied cell biological principles to address both issues. The resulting improved secretion signal confers dramatic benefits.

11:35 Titer Estimation for Quality Control (TEQC) Method: A Practical Approach for Optimal Production of Protein Complexes Using the Baculovirus Expression Vector System

Takagi_YuichiroYuichiro Takagi, PhD, Associate Professor, Department of Biochemistry and Molecular Biology, Indiana University School of Medicine

The baculovirus expression vector system (BEVS) is becoming the method of choice for expression of many eukaryotic proteins and protein complexes. However, what influences the overall production of proteins or protein complexes remains largely unclear. We developed the Titer Estimation for Quality Control (TEQC) method, which enables researchers to quantitatively optimize protein expressions utilizing BEVS in a highly reproducible fashion.

12:05 pm BryoTechnology: Large-Scale GMP-Manufacturing of Glyco-Designed Proteins with Moss

Schaaf_AndreasAndreas Schaaf, PhD, CSO, Greenovation Biotech GmbH

BryoTechnology, i.e., moss-based production of biopharmaceuticals, has evolved into a GMP manufacturing technology with products already in clinical development. While leveraging the mosses advantages, comparability to mammalian cell-based technologies was a priority in process development. Today’s moss process relies on the latest single-use technologies and follows the established routines of mammalian cell-based production. Thus, moss-based production fits easily into existing cleanroom environments and offers rapid changeover and flexible configuration.

12:35 End of Optimizing Protein Expression


5:45-8:45 pm Recommended Short Course*

SC15: Transient Protein Production in Mammalian Cells

Richard Altman, MS, Scientist, Protein Technologies, Amgen

Henry C. Chiou, PhD, Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

Dominic Esposito, PhD, Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research

 

*Separate registration required.

* 주최측 사정에 따라 사전 예고없이 프로그램이 변경될 수 있습니다.

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