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Protein expression used for the large-scale manufacturing of recombinant protein therapeutics and the production of target protein for drug screening and has become central to the biotechnology and pharmaceutical drug discovery and development process. Other applications of protein expression for both structural studies and protein interactions have grown with the development of structural genomic and proteomic research. However, as more genomic information has become available the logistics of protein expression have become more problematic. This Conference will examine the issues faced by scientists involved in maximising the efficiency of protein expression and overcoming the hurdles involved in purification, providing a valuable opportunity for researchers to get together to discuss emerging technologies and both practical and theoretical ideas.

The event will include a focus on some of the newer expression systems, the production of insoluble proteins for structure-function analysis and the production of hard to express proteins. The Conference will look to assess platform technologies and practical strategies in order to work towards highly-efficient, low background, high-throughput expression systems at low cost and ease of scale up.

A UNIQUE OPPORTUNITY TO HEAR FROM LEADING EXPERTS INCLUDING:
· Dr Jean-Dominique Guitton, Head, Lead Discovery Technologies, Aventis
· Dr Barbara Enenkel, Head, Molecular Biology, Process Science Department, Boehringer-Ingelheim
· Dr Ajith Kamath, Manager, Protein Science, Pfizer
· Dr Stefan Schmidt, Team Leader, Protein Science, AstraZeneca
· Dr Lukas Leder, Laboratory Head, Protein Expression & Purification, Novartis
· Dr Bernd Gerhartz, Laboratory Head, Protease Platform, Novartis
· Dr Angelo Gunasekera, Group Leader, Cancer Exploratory Biology, Abbott Laboratories
· Dr Tauseef Butt, President & Vice President, Research & Development, LifeSensors
· Dr Kenneth Lundstrom, Chief Scientific Officer, BioXtal

BENEFITS OF ATTENDING:
· OVERCOMING THE BOTTLENECKS: Optimise your strategies to overcome the difficulties presented by structural genomics
· PROTEIN EXPRESSION SYSTEMS: Evaluate the systems being used by industry leaders
· ENHANCING RECOMBINANT PROTEIN PRODUCTION: Identify and assess the promising new techniques being used
· EXPRESSING ‘DIFFICULT’ PROTEINS: Discover where the progress is being made
· NETWORK WITH KEY EXPERTS: Discuss and exchange ideas with other leaders in the field

Conference programme

8:30 Registration and Coffee

9:00 Chairman's Opening Remarks

Dr Tauseef Butt

Dr Tauseef Butt, President & Vice President, Research & Development, LifeSensors

9:10 TOWARDS INDUSTRIALISATION OF 3D DETERMINATION

Dr Jean-Dominique Guitton

Dr Jean-Dominique Guitton, Head, Lead Discovery Technologies, Aventis

  • Structural genomics: from initial dreams to reality
  • Impact of 3D structures on drug discovery: from target to EDC
  • From cDNA to structure: is protein production still the main bottleneck?
  • Automation, parellelisation, protein engineering: what are the possible solutions to overcome bottlenecks?
  • 9:40 PROTEINS FOR DRUG DISCOVERY EFFORTS

    Dr Ajith Kamath

    Dr Ajith Kamath, Manager, Protein Science, Pfizer

  • Different systems available for protein expression
  • Advantages of the baculovirus system
  • Reagents generation for high-throughput screening
  • Approaches used for producing proteins for crystallographic studies
  • 10:20 HIGH-THROUGHPUT PROTEIN EXPRESSION STUDIES

    Dr Angelo Gunasekera

    Dr Angelo Gunasekera, Group Leader, Cancer Exploratory Biology, Abbott Laboratories

  • Simple, highly efficient ligation-based cloning methods
  • Vectors with zero-background
  • PAGE-less solubility screen
  • Automated purification system
  • siRNA-based target validation
  • 11:00 Morning Coffee

    11:20 AN INTEGRATED CONCEPT FOR MAMMALIAN CELL LINE AND UPSTREAM DEVELOPMENT

    Dr Barbara Enenkel

    Dr Barbara Enenkel, Head, Molecular Biology, Process Science Department, Boehringer-Ingelheim

  • Platform technologies
  • Genetic engineering
  • Fast track cell line development
  • Process optimisation
  • Product quality
  • Scale-up and compatibility
  • 12:00 EXPRESSION, PURIFICATION AND FOLDING OF HUMAN PROTEINS - AUTOMATION AND STANDARDISATION

    Dr Konrad Büssow

    Dr Konrad Büssow, Laboratory Head, Max Planck Institute for Molecular Genetics/Protein Structure Factory

  • Cloning and availability of human full-length cDNA clones
  • Selection of target proteins for structural analysis
  • Expression of human proteins in E. coli
  • Parallel, automated protein purification
  • Automated protein folding screen for inclusion body proteins
  • 12:40 Networking Lunch

    13:40 PER.C6(TM) AS A PLATFORM FOR MANUFACTURING OF MONOCLONAL ANTIBODIES

    Dr Richard van Rijnsoever

    Dr Richard van Rijnsoever, Manager, Business Development, Crucell

  • Background of the cell line
  • Selecting high expressing cell lines
  • Expression levels of monoclonal antibodies
  • Feed strategy
  • Scaling of the process
  • Quality of the monoclonal antibodies
  • 14:20 AUTOMATED PROTEIN PURIFICATION AND UP-SCALING STRATEGIES

    Dr Stefan Schmidt

    Dr Stefan Schmidt, Team Leader, Protein Science, AstraZeneca

  • Current status of purification automation at AstraZeneca
  • Tackling homogenisation
  • Solubility issues
  • Addressing analytical bottlenecks
  • Strategies for up-scaling automated purification (5P)
  • Further perspectives
  • 15:00 Afternoon Tea

    15:20 SEPARATION TECHNOLOGIES APPLIED TO PROTEIN DISCOVERY, INTERACTION AND PURIFICATION

    Dr Egisto Boschetti

    Dr Egisto Boschetti, Vice President, Research, Ciphergen

  • Why and how to separate proteins
  • Differences in phenotypes can be evidenced by preliminary proteome fractionation
  • Biomarkers are related to the modification of interactors and evidenced by ProteinChip® Technology
  • Purification is necessary all along the process or proteomic investigation
  • Discovery of new proteins induces specific purification procedures starting from very small samples
  • Prediction of separation condition as a critical point to overcome. This is possible with SELDI-MS technology
  • 16:00 INCREASING THE SOLUBILITY OF RECOMBINANT PROTEINS

    Dr Arie Geerlof

    Dr Arie Geerlof, Head, Protein Expression & Purification, European Molecular Biology Laboratory (EMBL)

  • The role of bacterial chaperones in protein folding
  • Co-expression of different combinations of bacterial chaperones
  • Protein de-aggregation by bacterial chaperones
  • Small scale expression screening
  • Increased solubility for more than 70% of the tested target proteins
  • Up to 42-fold more soluble protein obtained
  • 16:40 THE EXPRESSIONFACTORY(™) - A PLATFORM TECHNOLOGY FOR SYSTEMATIC, AUTOMATED AND HIGH THROUGHPUT PROTEIN EXPRESSION AND PURIFICATION

    Dr Grant Cameron

    Dr Grant Cameron, Commercial Director, NextGen Sciences

  • This presentation will discuss NextGen Sciences' expressionfactory, which fully automates protein expression
  • The system integrates a unique combination of biology, hardware and software tools, enabling the user to produce highly purified proteins, automatically
  • Expression vector construction, protein expression and subsequent purification are fully automated, resulting in the systematic, parallel production of many hundreds of proteins with minimum input and hands-on time.
  • A range of unique expression vector systems will be discussed which incorporate a series of fusion partners, enabling optimal soluble protein expression, together with affinity tags for subsequent purification.
  • It is possible to start with PCR products and cDNA clones and plan, schedule, automate and track the whole process of sub-cloning, expression and purification of hundreds of proteins in parallel.
  • The advanced capabilities of the system will be demonstrated with examples including the automated expression of human p53 protein
  • 17:20 A versatile cell-free expression system - The Rapid Translation System offers new options for the rapid expression of proteins

    Dr Sabine Wisemann

    Dr Sabine Wisemann, Scientific Product Support, Roche Diagnostics

  • Flexible protein expression workflow including bioinformatics based sequence optimization
  • High initial success rate in protein expression using the wheat germ lysate
  • Truly scalable high yield E.coli lysate
  • 18:00 Chairman's Closing Remarks and Close of Day One

    8:30 Re-registration and Coffee

    9:00 Chairman's Opening Remarks

    Dr Kenneth Lundstrom

    Dr Kenneth Lundstrom, Chief Scientific Officer, BioXtal

    9:10 INDUSTRIAL PROTEIN REFOLDING

    Dr Laurent Vuillard

    Dr Laurent Vuillard, Executive Head, Operations, Avidis

  • Why in vitro refolding?
  • Additives and folding screen
  • Screening without activity readout
  • Small scale to scaleup
  • Success rate?
  • 9:40 THE PROTEIN AS A VARIABLE IN PROTEIN CRYSTALLISATION

    Dr Glenn Dale

    Dr Glenn Dale, Head, Biochemistry, Morphochem

  • Screening and modifications
  • Point mutations
  • Insertions and deletions
  • Fusion proteins
  • Perspectives
  • 10:20 ANALYTICAL METHODS FOR PROTEIN CHARACTERIZATION

    Dr Stefan Geschwinder

    Dr Stefan Geschwinder, Associate Principal Scientist, AstraZeneca

  • Engineering proteins for structural studies and the use of the analytical toolbox
  • Demands on physical, chemical and functional characterisation methods to check protein function, stability and quality
  • The analytical toolbox: focusing on thermodynamic and spectroscopic methods for protein characterisation
  • Moving the bottleneck: consequences of HT cloning and expression capabilities on protein characterisation and how we cope with that
  • 11:00 Morning Coffee

    11:20 MULTIPARALLEL PROTEIN EXPRESSION ANALYSIS

    Dr Lukas Leder

    Dr Lukas Leder, Laboratory Head, Protein Expression & Purification, Novartis

  • Expression of different constructs with a target protein under various conditions
  • Lysis of cells and separation into soluble and non-soluble extracts done on liquid handling workstation
  • Fast and homogeneous quantification of target proteins by reconstitution of active ribonuclease S with added S-protein and internal S-tag
  • Cleavage of an oligonucleotide substrate with a fluorescent dye and quencher pair yields a high fluorescent signal
  • Some effects on sensitivity caused by adjacent fusion tags
  • Results highly comparable to quantification done by SDS-PAGE
  • 12:00 BIOPHYSICAL APPROACHES TO TACKLE ‘DIFFICULT’ PROTEINS

    Prof Peter Henderson

    Prof Peter Henderson, Professor, Biochemistry & Molecular Biology, University of Leeds

  • Amplified expression, purification and reconstitution
  • CD and FTIR spectrometry
  • Fluorimetry
  • Solid state and solution state NMR
  • 2D crystallisation and electron diffraction
  • 3D crystallisation and X-ray diffraction
  • 12:40 Networking Lunch

    13:40 SUMO GENE FUSIONS

    Dr Tauseef Butt

    Dr Tauseef Butt, President & Vice President, Research & Development, LifeSensors

  • The role of SUMO and Ubiquitin in protein trafficking and modification
  • Using a SUMO fusion system to overcome the problems faced by traditional gene-fusion technologies
  • Enhancing the expression of under-expressed or non-expressed proteins
  • Improving solubility and proper folding of insoluble proteins in prokaryotes and eukaryotes
  • Application of the system and development of newer systems to express and purify a wide variety of proteins in prokaryotes and eukaryotes
  • 14:20 EXPRESSION OF GPCRs - ART OR SCIENCE?

    Dr Kenneth Lundstrom

    Dr Kenneth Lundstrom, Chief Scientific Officer, BioXtal

  • Expression of a large number (100) of GPCRs
  • 3 expression systems, based on bacterial, yeast and viral vectors
  • Expression of a large number of targets at 1 mg/L level or more
  • Refolding / solubilisation of recombinant GPCRs
  • Purification and crystallisation of GPCRs
  • 15:00 NUCLEAR RECEPTORS FOR DRUG DESIGN

    Dr Mats Carlquist

    Dr Mats Carlquist, Head, Structural Biology, Karo Bio

  • Design of target protein
  • Fusion tags
  • Solubility
  • Chemical modification
  • Mutations
  • 15:40 Afternoon Tea

    16:00 ASPARTYL PROTEASES

    Dr Bernt Gerhartz

    Dr Bernt Gerhartz, Laboratory Head, Protease Platform, Novartis

  • Comparing different expression systems on specified examples
  • Comparing different members of the aspartyl protease family in one expression system
  • Comparing refolding of different family members
  • Addressing special problems of activation, autodegradation, etc
  • 16:40 THE GATEWAY SYSTEM: FACILITATING RECOMBINANT PROTEIN EXPRESSION BY ALLOWING RAPID GENE CLONING AND TRANSFER

    Harry Yim

    Harry Yim, Staff Scientist, Invitrogen

  • Recombinational Cloning: Learning from viruses
  • The Ultimate ORF collection: A fully sequence validated, Gateway compatible Human and Mouse Open Reading Frame collection.
  • The Gateway Open Architecture
  • Expression tools: Gateway compatible structural and functional proteomics systems
  • 17:20 Chairman’s Closing Remarks and Close of Conference

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