PEGS Europe 2010 | Day 1 | Day 2
Despite the great strides made in the field of protein expression and purification, there still remain a great number of “difficult” proteins which continue to challenge existing methods. However, new technologies are emerging which hold the promise of improving yields, simplifying isolation and characterization and advancing the capabilities of protein engineering, particularly for these difficult to express proteins. This conference will present the new technologies and methodologies, as well as cutting-edge applications of this break-through science.
WEDNESDAY, 6 OCTOBER
13:00 Conference Registration
14:00 Chairperson’s Remarks
Kenneth Lundstrom, Ph.D., CEO, PanTherapeutics
14:05 Cell-Free Production of Difficult Pharmaceutical Targets: Case Studies of G-Protein Coupled Receptors and Alzheimer’s Disease Related Proteins
Frank Bernhard, Ph.D., Centre for Biomolecular Magnetic Resonance, Institute for Biophysical Chemistry, University of Frankfurt/Main
Cell-free expression eliminates major bottlenecks in the synthesis of membrane proteins and allows their production in completely new modes. We exemplify protocol development strategies for the efficient production of GPCRs in individual cell-free systems. We further demonstrate the high quality production of the Endothelin receptom, the major player in blood pressure regulation, and we present structural data of cell-free produced subunits of the y-secretase complex.
14:35 Semliki Forest Virus Vectors: Versatile Tools for Protein Production
Kenneth Lundstrom, Ph.D., CEO, PanTherapeutics
Recombinant protein production is an essential part of modern biotechnology. Semliki Forest virus (SFV) vectors have been applied for protein production in drug development and structural biology and in neuroscience and gene therapy. SFV is particularly useful for the expression of difficult proteins such as membrane receptors. Rapid high-titer virus production, broad host range, high levels of transient gene expression are attractive features of SFV.
15:05 High-Level Production and Characterization of a G-Protein Coupled Receptor Signaling Complex
Stephen Marino, Ph.D., Department of Molecular Membrane Biology, Max Planck Institute of Biophysics
The presentation describes the combination of factors permitting the successful overproduction of a physiologically relevant G protein-coupled receptor (GPCR) complex. An evaluation of these factors (generalizable for other receptors), their advantages and disadvantages, as well as demonstration of the functionality of the resulting receptor species, will be discussed.
15:35 Refreshment Break
16:00 Advancing Synthetic Gene Design
Claes Gustafsson, Ph.D., Vice President, Marketing and Sales, DNA2.0
Gene synthesis offers immense flexibility in the tailoring of genes for practical uses. Capturing the value of this flexibility, however, is greatly limited by lack of understanding of the interactions between gene sequence features and host expression systems. DNA2.0 has developed a novel approach to interrogate the gene design preferences of expression hosts to maximize production from synthetic genes. Applications of this approach for a number of target proteins in several different host organisms will be discussed.
16:15 An Accelerated Protein Stability Testing Assay For Optimizing Purification Steps and Storage Conditions
Wayne F. Patton, Ph.D., Chief Scientific Officer, Enzo Life Sciences
Aggregation tendency is often studied by exposing a protein solution to extreme conditions of temperature, pH, humidity, and light, referred to as forced photodegradation studies. Proteins degraded in this manner can reflect the degradation pathway(s) experienced during the product's lifetime. A rapid high-throughput fluorescence-based assay for assessing stability of monoclonal antibodies and recombinant therapeutic proteins is described. The ProteoStat® Thermal Shift Assay is performed using a temperature-regulated fluorimeter or a real-time PCR instrument. The assay is suitable for screening for protein stability as a function of pH, ionic strength, and concentration and the analysis of ligand binding.
16:30 Single-Chain Constructs of the Active Liver X Receptor for Enhanced Solubility and Improved X-Ray Diffraction
Zhongren Wu, Research Fellow, Biochemistry, Vitae Pharmaceuticals
Liver X receptors (LXRs) are members of the nuclear receptor superfamily. We have developed novel, single-chain expression constructs that fuse a co-activator peptide to the LBD domain of LXRs through a flexible linker. The proteins are simple to purify with high yields. High quality co-crystal structures have been obtained with a number of ligands using this approach, including a LXRα homodimer complexed with T0901317 at 1.7Å.
17:00 EasyProt: An Innovative Protein Expression System Based on the Secretion System Type III of Pseudomonas aeruginosa
Audrey Le Gouellec, Ph.D., Assistante Hospitalo-Universitaire, HumProTher Laboratory, TheREx-GREPI, TIMC-IMAG Laboratory, University of Joseph Fourier, UFR de Médecin
We present a technology to produce recombinant proteins using P. aeruginosa strain. The type III secretion system is used to secrete into the medium the heterologous proteins produced, thus facilitating their purification and functional analysis. The secretion of proteins into the oxidative extracellular medium could favor natural folding and disulfide bridge formation. We used this technology to screen vaccine antigen for anti-tumoral immunotherapy.
18:30 – 21:00 BIOTECHNICA Night: Beer Hall, Full Dinner Reception, Live Band
(Please register to reserve your complimentary ticket ahead of time. No tickets will be available on-site.)