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2017年11月13-14日

TS1:タンパク質製造研究室の基本的な技術


Instructors:
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
Mario Lebendiker, Ph.D., Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology

This seminar is designed to introduce basic technologies, strategies and considerations in recombinant protein production in E. coli, insect and mammalian cells for multiple research and development applications. The seminar supplies a basic toolbox for management of multiple and diverse projects.

Brief Description:

  • Overview of recombinant protein production: initiating a project, design and options for various downstream applications, requirements from customer/collaborator, and matching expectations.
  • Introduction to expression systems; covering the differences between the standard expression systems: E. coli, insect cells and mammalian cells, in protein quality, quantity and downstream applications, cost considerations, implementation time, required expertise and more.
  • Basic principles in affinity chromatography, ion exchange, hydrophobic exchange, size exclusion and mixed mode chromatography. Protein purification strategies: input for purification protocol development, guidelines for protein purification, selection and combination of purification techniques.
  • Connection between expression and protein quality, dealing with prone-to-aggregate proteins, selection and combination of purification techniques, major requirements for purification of proteins for structural, biophysical and biochemical studies, minimal quality control parameters and quality control workflow.
  • Troubleshooting and case studies.

Who should attend the training seminar?
The seminar is designed for researchers, lab managers, graduate students, postdocs, technicians and engineers wishing to expand their knowledge and implement basic and advanced technologies in recombinant protein production in their work.

Detailed Agenda

MONDAY, 13 NOVEMBER

12:00 Registration

13:40 Training Sessions Module One: Overview of Recombinant Protein Production in Cellular Systems and Project Initiation

  • Initiating a project in consideration of downstream applications and target protein characteristics
  • E. coli-based protein production: overview on vectors and hosts
  • Baculovirus-based expression in insect cells: cell lines, growth conditions, and commercial systems overview
  • Mammalian expression systems and cell-based assays

15:50 Refreshment Break in the Exhibit Hall with Poster Viewing

16:50 Training Sessions Module Two: Basic Principles in Chromatography

  • Protein purification: main stages, flowchart
  • Ion exchange, hydrophobic exchange, mixed mode, affinity and gel filtration chromatography: advantages and disadvantages
  • Basis for selectivity, operational considerations, determinations of start conditions, parameters for optimization

18:25 Welcome Reception in the Exhibit Hall with Poster Viewing

19:25 End of Day One


TUESDAY, 14 NOVEMBER

07:45 Morning Coffee

08:30 Training Sessions Module Three: Selecting Strategies for Protein Purification

  • Input for purification protocol development
  • Selection and combination of purification techniques, connection between expression and protein quality
  • Major requirements for purification of proteins for structural studies: crystallography, NMR, etc.

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Training Sessions Module Four: Troubleshooting and Case Studies in Protein Expression Procedures

  • Heat shock procedures for enhancing solubility in E. coli
  • Exploiting the potential of E. coli systems using quick screens
  • Secreted and membrane proteins in insect cells

12:45 Lunch

13:30 Dessert Break in the Exhibit Hall with Poster Viewing

14:00 Training Sessions Module Five: Managing Multiple Projects in a Non-Automated Environment

  • Service strategies and structure of protein production lab
  • Training facility personnel vs. training students and users
  • Prioritizing projects
  • Psychological aspects

16:05 Refreshment Break in the Exhibit Hall with Poster Viewing

16:45 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 seminar's panel of experts 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:

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


Panelists:

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

Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

Dominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research

Mario Lebendiker, Ph.D., Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem

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

18:15 End of Training Seminar 1



Instructor Biographies:

Tsafi_DanieliTsafi 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
Tsafi Danieli is the head of the Protein Expression Core Facility at the Wolfson Centre for Applied Structural Biology at the Hebrew University of Jerusalem, Israel. The core facility functions as a biotechnology research center for the development and implementation of novel technologies in recombinant DNA, molecular biology and protein production. It also operates as a training centre supporting individual research projects and conducting workshops and courses for scientists from academic institutes and from the biomed & biotech industry. Dr. Danieli is also the founder and director of "BioGiv", a specialised university-based center for promoting and supporting entrepreneurs and inventors to translate ideas into new products and bring new technologies to market, while using the infrastructure of the Hebrew University in Jerusalem. Dr. Danieli holds a BSc in biology and MSc in human genetics from Tel Aviv University, and a Ph.D. in molecular virology from the University of California, San Francisco Pharmacology Department, in collaboration with the Biochemistry Department at Tel Aviv University.

Mario_LebendikerMario Lebendiker, Ph.D., Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem
Dr. Mario Lebendiker is in charge of the Protein Purification Facility at the Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem. He is actively involved in many collaborations for structural and biochemical studies within the Hebrew University, others Universities in Israel, as well as with biotech and pharmaceutical companies. Dr. Lebendiker received a Ph.D. in Biochemistry in 1982 from the Animal Virology Center (CEVAN), in Buenos Aires University, Argentina. Together with many other laboratories, he found the Protein Production and Purification Partnership in Europe (P4EU) network, a platform for the exchange of information, knowhow and materials between core facility labs in the field of protein expression and purification.

Panelist Biographies:

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

Rich Altman has 29 years of experience working in the pharmaceutical industry. In 2016, he joined the Protein Technologies Mammalian Expression group at Amgen San Francisco, supporting biologics drug development. Prior to Amgen, he worked for several pharmaceutical companies on the cloning, expression, purification and characterization of recombinant proteins. This work supported both small-molecule high-throughput screening and protein therapeutic efforts. He received his MS degree from the University of Pittsburgh School of Medicine in the Department of Molecular Biology and Biochemistry.

Chiou HenryHenry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

Dr. Chiou has over 15 years of experience working on development of mammalian protein expression systems and transfection reagents. For the past nine years he has worked as a principal scientist and now in product development and portfolio management for Life Technologies, now part of Thermo Fisher Scientific. He has led the development efforts on a number of products, including the FreeStyle™ transient expression systems, Lipofectamine™ LTX and Lipofectamine™ RNAiMAX. Prior to Life Technologies, he worked for several biotech companies on biotherapeutic gene delivery systems. Dr. Chiou received his doctorate from Harvard University and completed a postdoctoral fellowship at the University of Pennsylvania.

Esposito DominicDominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research

Dr. Esposito is currently the Director of the Protein Expression Laboratory (PEL) at the Frederick National Laboratory for Cancer Research. The PEL is currently focused on producing Ras and Ras-related proteins for the National Cancer Institute's RAS Initiative. These proteins are being used for a wide-ranging attack on mutant Ras genes including structural biology, biophysics and development of screens and assays for drug discovery. In addition, the PEL still assists in the generation of proteins of interest to investigators in the intramural program of the NCI, and invents and develops new technologies for protein expression and production. Dr. Esposito received his BA in Chemistry at La Salle University in Philadelphia, and his Ph.D. in Biochemistry at the Johns Hopkins University Bloomberg School. Dr. Esposito previously worked for Life Technologies, where he helped to develop the Gateway recombinational cloning system.

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

Bjorn Voldborg is a veteran within the field of recombinant protein expression, with more than 20 years of experience from both academic and industrial settings, working with recombinant protein expression, both as part of the development of cell-based assays, as well as producing recombinant proteins for drug discovery for academic and industrial research. At the Institute of Molecular Biology at the University of Copenhagen Bjorn developed cell based assays to measure activity of drosophila receptors. At the Danish biotech company Pharmexa A/S, Bjorn was teamleader in the molecular biology group, responsible for molecular cloning and initial expression test of potential proteinbased drug candidates. From this he went to the NNF Center for Protein Research at the University of Copenhagen as Head of the Protein Production Unit, and since 2012 Bjorn has been heading the CHO Cell Line engineering project at the NNF Center for Biosustainability at the Technical University of Denmark, a project dedicated to the engineering of improved protein production cell factories.



Training Seminar Information

Each CHI Training Seminar offers 1.5 days of instruction with start and stop times for each day shown above and on the Event-at-a-Glance published in the onsite Program & Event Guide. Training Seminars will include morning and afternoon refreshment breaks, as applicable, and lunch will be provided to all registered attendees on the full day of the class.

Each person registered specifically for the training seminar will be provided with a hard copy handbook for the seminar in which they are registered. A limited number of additional handbooks will be available for other delegates who wish to attend the seminar, but after these have been distributed, no additional books will be available.

Though CHI encourages track hopping between conference programs, we ask that Training Seminars not be disturbed once they have begun. In the interest of maintaining the highest quality learning environment for Training Seminar attendees, and because Seminars are conducted differently than conference programming, we ask that attendees commit to attending the entire program, and not engage in track hopping, as to not disturb the hands-on style instruction being offered to the other participants.


* 不測の事態により、事前の予告なしにプログラムが変更される場合があります。

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