2018 Archived Content
OVERVIEW | DOWNLOAD BROCHURE | SPEAKERS | SHORT COURSES

TS6A: BASIC TECHNOLOGIES IN A PROTEIN PRODUCTION LAB
MONDAY 12 NOVEMBER AND TUESDAY 13 NOVEMBER
Day 1: 13:30 - 19:30 | Day 2: 08:30 - 18:30

Room Location: Room 5B

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.

Instructors:

Tsafi Danieli, PhD, 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, PhD, Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem

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, 12 NOVEMBER

12:00 Registration

13:30 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

15:45 Networking Refreshment Break

16:15 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

18:20 Welcome Reception in the Exhibit Hall with Poster Viewing

19:30 End of Day One


TUESDAY, 13 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.

10:30 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Training Sessions Module Four, Part 1: Troubleshooting and Case Studies in Protein Expression Procedures

  • Aggregations truncations and low productivity in E. coli
  • Exploiting the potential of E. coli systems in a limited time frame
  • Common problems in baculovirus-based expression in insect cells
  • Standardization and operating procedures in mammalian cells

12:45 Lunch Provided

13:45 Dessert Break in the Exhibit Hall with Poster Viewing

14:15 Training Sessions Module Four, Part 2: Standardization and Operating Procedures in Mammalian Cells

15:05 Training Sessions Module Five: Managing Multiple Projects in a Non-Automated Environment

  • Service strategies and structure of protein production lab
  • Production services vs. training students and users
  • Prioritizing projects
  • Psychological aspects

16:20 Refreshment Break in the Exhibit Hall with Poster Viewing

17:00 Instructor Wrap-Up

17:30 CLOSING PANEL DISCUSSION: Tools for Expanding the Protein Engineering and Production Toolbox

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, PhD, 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, Protein Technologies, Amgen

Nicola Burgess-Brown, PhD, Principal Investigator, Biotechnology, Structural Genomics Consortium (SGC), University of Oxford

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

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

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

Jonathan Zmuda, PhD, Director, Cell Biology, Thermo Fisher Scientific

18:30 End of Training Seminar 6A

Instructor Biographies:

Danieli_TsafiTsafi Danieli, PhD, 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 and training center for the development and implementation of novel technologies in molecular biology and protein production. Dr. Danieli is also the founder and director of “BioGiv Excubator”, a specialized university-based center for promoting and supporting early-stage startup companies 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 PhD in molecular virology from the Biochemistry Department at Tel Aviv University in collaboration with the University of California, San Francisco Pharmacology Department.

Lebendiker_MarioMario Lebendiker, PhD, 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 PhD 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, Protein Technologies, Amgen

Rich Altman has 30 years of experience working in the pharmaceutical industry. In early 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.

Burgess-Brown_NicolaNicola Burgess-Brown, PhD, Principal Investigator, Biotechnology, Structural Genomics Consortium (SGC), University of Oxford

Nicola Burgess-Brown is the Principal Investigator of the Biotechnology Group at the SGC, responsible for managing all biotech research for the Oxford site. Nicola’s team optimises the high-throughput screening processes from cloning to expression and purification of human proteins for structural and functional studies. The group collaborates and interacts closely with the other SGC teams, to develop methods for increasing protein expression, parallel processing and increasing throughput for soluble and integral membrane proteins (IMPs) involved in human disease. Nicola obtained a First Class degree in Applied Biochemical Sciences from the University of Ulster in 1997 and spent the following year working as a molecular biologist for SmithKline Beecham. She received her PhD in Molecular Microbiology at the University of Nottingham in 2001 and then moved back to industry to work on high-throughput cloning and validation of therapeutic cancer antigens for Oxford Glycosciences and subsequently Celltech R&D.

Esposito_DominicDominic Esposito, PhD, 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 providing production of Ras and Ras-related proteins for the National Cancer Institute’s RAS Initiative. These proteins are being used for a wide-ranging attack on Ras biology 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 PhD 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.

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

Bjørn Voldborg has worked within the field of recombinant protein expression, for more than 20 years with experience from both academic and industrial settings, 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 Bjørn developed cell based assays to measure activity of drosophila receptors. At the Danish biotech company Pharmexa A/S, Bjørn 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 Bjørn 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 cell factories for protein production.

Vzmuda jonathan Jonathan Zmuda, PhD, Director, Cell Biology, Thermo Fisher Scientific

Jonathan Zmuda, PhD is a Director of Cell Biology in the Life Sciences Solutions Group at Thermo Fisher Scientific in Frederick, MD (USA). Dr. Zmuda leads a team of scientists dedicated to developing new technologies for cell biology applications including protein expression, advanced cell culture and rare cell analysis. Dr. Zmuda received his PhD in Cell Biology from the University of Maryland, College Park and his undergraduate degree from Dickinson College in Carlisle, PA.