Archived Content

Sixth Annual Protein Expression

 

Part Two: Solving Difficult Protein Problems


Day 1 | Day 2 
Part One | Download Brochure 

 

Wednesday 7 November

12:30 – 14:00 Conference Registration for Part Two

14:00 Chairperson’s Remarks

M. Raafat El-Gewely, Ph.D. Professor, Institute of Medical Biology, University of Tromsø

Membrane Proteins

»Keynote PRESENTATION

14:05 Nanobodies as Tools for the Structural and Functional Investigation of GPCR Transmembrane Signaling

Jan SteyaertJan Steyaert, Ph.D., Executive Director, Department of Molecular and Cellular Interactions, Vlaams Instituutvoor Biotechnologie & Structural Biology Brussels, Vrije Universiteit Brussel

We generated Nanobodies that stabilize transient functional conformations of the human b2 adrenergic receptor. Some Nanobodies that faithfully mimic G protein binding were used to crystallize active agonist-bound states of this GPCR. Other nanobodies that stabilize the b2AR•Gs complex were instrumental to purify and obtain the crystal structure of this complex, providing the first view of transmembrane signaling by a GPCR.


14:35 Optimization of Expression and Purification of the Feline and Primate Foamy Virus Transmembrane Envelope Proteins Using a 96 Deep Well Screen

Michael MühleMichael Mühle, Ph.D., Researcher, Robert Koch Institute

High level overexpression of a recombinant target protein in E. coli is a prerequisite to minimize efforts in subsequent large scale purification steps. We developed a 96 well based screening method for rapid detection of suitable expression conditions and applied it for the production of two viral transmembrane envelope proteins to quickly determine conditions for an about 20-fold increased production.

BAC15:05 CaptureSelect: Unique Affinity Products for Purification and Detection of All Antibody Formats

Frank Detmers, Ph.D., Director, Ligand Application, BAC BV

This presentation will focus on CaptureSelect IgG-CH1, a novel affinity resin, using a ligand directed towards the CH1 domain of antibodies and antibody fragments, offering a true platform for purification of all human IgG’s and Fab fragments thereof, irrespective of subclass and without co-purification of free light chains. Analytical assays based on HPLC column formats and label free detection systems for QC will also be presented.

15:20 Sponsored Presentation

(Opportunity available, please contact Carol Dinerstein, Dinerstein@healthtech.com)

15:35 Refreshment Break in the Exhibit Hall with Poster Viewing

16:15 Predictable Protein Expression in Mammalian Cells by Exploitation of Defined Chromosomal Loci

Dagmar Wirth, Ph.D., Department of Gene Regulation and Differentiation, HZI- Helmholtz Centre for Infection Research

The expression levels of transgenes in mammalian cells are affected by the impact of neighbouring chromosomal elements. To achieve predictable protein expression we exploited methods for targeting expression cassettes to specific chromosomal sites via recombinase mediated cassette exchange or employing bacterial artificial chromosomes, respectively. We show that defined combinations of integration sites and vector designs are required for rational exploitation of given chromosomal sites.

16:45 Improving Protein Stability and Function through Disulfide Engineering: A Computational Approach

Alan DombkowskiAlan Dombkowski, Ph.D., Assistant Professor, Division of Clinical Pharmacology and Toxicology, Department of Pediatrics, Wayne State University School of Medicine

Protein stability and function can be enhanced in a wide range of biomedical and industrial applications through disulfide bond engineering. Computational methods have proven to be effective for designing strategically placed disulfides. We will discuss structural considerations for successful use of this technology, recent advances in our computational method for rational disulfide design, and applications.

17:15 Problem Solving Roundtable Discussions

Table 7: Nanobodies as Tools for Structural Studies of Membrane Proteins

Moderator: Jan Steyaert, Ph.D., Executive Director, Department of Molecular and Cellular Interactions, Vlaams Instituutvoor Biotechnologie & Structural Biology Brussels, Vrije Universiteit Brussel

Table 8: Predicting Protein Expression in Mammalian Cells Using Defined Chromosomal Loci

Moderator: Dagmar Wirth, Ph.D., Department of Gene Regulation and Differentiation, HZI- Helmholtz Centre for Infection Research

Table 9: Cell Free Expression of Mammalian Proteins: Strategies for Success

Moderator: Frank Bernhard, Ph.D., Centre for Biomolecular Magnetic Resonance, Institute for Biophysical Chemistry, University ofFrankfurt/Main

Table 10: Using Bacterial Artificial Chromosomes to Enhance Protein Expression in Mammalian Cell Lines

Moderator: Emilio Casanova, Ph.D., Researcher, Ludwig Boltzmann Institute for Cancer Research, Vienna

Table 11: Choosing the Right Host for Expression of Membrane Proteins

Moderator: Annie Barrand-Frelet, Ph.D., CNRS, CEA-France


18:15 – 19:15 Reception in the Exhibit Hall with Poster Viewing



Day 1 | Day 2 
Part One | Download Brochure