Day 1 | Day 2 | Download Brochure
Although most scientists would argue that the science of protein expression is a mature one, there still exist many issues with yields, glycosylation, folding, inclusion bodies, and so on. There is still much that can be learned to overcome these challenges. New hosts are emerging that will provide solutions to these ongoing problems. In addition, new methods and technologies continue to provide improved yield and stability of proteins in the existing hosts. Cambridge Healthtech Institute’s 13th Annual “Enhancing Host Expression” (formerly “Protein Expression”) conference will present the cutting-edge methods and technologies that improve the outcomes in existing host systems, as well as new and novel hosts that will become the tools for the 21st Century protein expression scientist.
Recommended Pre-Conference Short Course *
Sunday, January 10
(SC4) Specialized Protein Expression Systems - An Overview
* Separate registration required
Monday, January 11
7:30 am - 8:45 am Registration and Morning Coffee
8:45 Chairperson’s Opening Remarks
Dominic Esposito, Ph.D., Principal Scientist, Protein Expression Laboratory, SAIC-Frederick, Inc.
9:00 KEYNOTE PRESENTATION
A Review of High Throughput Protein Production in Drug Discovery: Lessons Learned
Ian Hunt, Ph.D., Protein Structure Unit, Centre for Proteomic Chemistry, Novartis Institutes for BioMedical Research
The presentation will review some of the technologies and innovations that have been developed to accelerate the generation of recombinant proteins for the support of early drug discovery (HTS, biophysics & structural biology) within the pharmaceutical environment and will include some tricks of the trade and a look ahead at the future of protein production.
9:50 General and Adaptable High-Throughput Approaches for the Optimization of Protein Expression
Ian Fotheringham, Ph.D., President, Ingenza Ltd.
A novel screening system has been developed and demonstrated that allows for the high-throughput detection of genetic variants that exhibit increased expression levels. The system is highly tunable and versatile and relies on the fusion of the gene of interest to a reporter gene encoding an oxidase. The system is built on many years of experience of screening for oxidase activity and has been fully validated and used to identify specific codon optimized sequences from large libraries of variants.
10:20 Networking Coffee Break
10:45 Auditioning of CHO Host Cell Lines Using the Artificial Chromosome Expression (ACE) Technology
Malcolm L. Kennard, Ph.D., Kennard Biologic Consultants
The ACE System, based on targeted transfection through pre-engineered artificial chromosomes, was investigated as a technique to audition host cell lines. Clonal cell lines were generated from three CHO host cell lines (CHOK1SV, CHO-S and DG44) that had identical artificial chromosomes containing gene copies of a human monoclonal IgG1 antibody. The resulting large differences in antibody expression between the different CHO host cells were shown to be due to cell phenotype rather than genetic factors.
11:15 Anti-Apoptotic Factors Improve Productivity of Monoclonal Antibodies from CHO Cells
Mark Levenstein, Ph.D., Scientific Director, Biochemistry, CDI Bioscience
In batch bioreactors overpopulation, nutrient depletion, and waste accumulation create a hostile environment for in vitro cell culture, triggering apoptosis, cutting short the productive phase of a recombinant therapeutic production run. With access to more ATP individual cells produce more protein. An expanded endoplasmic reticulum facilitates synthesis and secretion of protein products into the culture medium. In CHO cells the combined effects of p16 and p21 result in a three-fold increase in protein productivity as measured in picograms per cell per day.
11:45 Using the Genome(s) to Optimize Secreted Heterologous Protein Expression
Thomas Chappell, Ph.D., Founder and Principal Scientist, BioGrammatics, Inc.
The recent publication of genome sequences from several Pichia pastoris strains and/or closely related species has provided the basis for optimization of protein expression in this widely used host system. Our focus has been on generating a panel of reporter strains where the stress response to a specific heterologous protein can be quantified in different genetic backgrounds. This output can then be used to choose specfic expression partners to optimize protein folding and transit through the secretory pathway.
12:15 pm Close of Morning Session
12:30 Luncheon Presentation
Focus on Your Freedom to Innovate: How Blue Sky’s BioProducts & Collaborative Product Development Alliances Can Help You
Scott Gridley, Ph.D., Head of Product Development, Blue Sky Biotech
1:15 Luncheon Presentation
Advanced Method of Cell Line Generation Using UCOE® Technology
Kim Mann, Sr. Research Scientst, Millipore
Joe Orlando, Sr. Research Scientist, Millipore
Anjali Verma, Research Scientist, Millipore
Isolation of highly-productive stable cell lines is the primary obstacle in the process of therapeutic protein production. Methods such as gene amplification and automated clone selection have been somewhat successful in overcoming this barrier. However, these methods do have drawbacks such as increased timelines, instability of expression and increased cost to the process.
UCOE (Ubiquitous Chromatin Opening Element) technology overcomes impediments of cell line development by producing numerous clones from a single transfection that are both highly-productive and stable. The studies presented here demonstrate that UCOE technology can help overcome obstacles for recombinant protein production.
2:00 Chairperson’s Remarks
Dominic Esposito, Ph.D., Principal Scientist, Protein Expression Laboratory, SAIC-Frederick, Inc.
2:35 High-Throughput Protein Expression Using Cell-Free Systems
Deb K. Chatterjee, Ph.D., Protein Expression Laboratory, SAIC-Frederick, Inc., National Cancer Institute
Traditionally, the cell-free transcription/translation system has been used mainly for analytical purpose to characterize gene products. However, recent developments made cell-free protein synthesis as an alternative to cell-based synthesis and solved many of problems associated with in vivo systems. Furthermore, this technology is readily amenable to automation. We will present examples for expressing recombinant proteins with high yield (mg quantities) using highly active S30 extract in home-made inexpensive dialysis method in a high-throughput manner.
3:05 Hijacking the Regulated Secretory Pathway of for High-Level Expression of Recombinant Proteins
Theodore Clark, Ph.D., Associate Professor, Microbiology and Immunology, Cornell University
The common pondwater ciliate, Tetrahymena thermophila, stores large amounts of protein in several thousand dense core secretory granules that can be triggered to discharge their contents to the extracellular space at will. The material secreted from cells takes the form of a proteinaceous gel that can be readily harvested by low speed centrifugation. When combined with ribosomal DNA vectors that replicate to ~18,000 copies per cell as palindromic minichromosmes, the regulated secretory pathway of Tetrahymena can be harnessed for high-level expression and streamlined purification of recombinant proteins in a process that is tantamount to affinity chromatography in vivo.
3:35 Pfēnex Expression TechnologyTM: The New Standard for Protein Expression
Diane Retallack, Ph.D., Director of Molecular Biology, Pfenex Inc.
The Pseudomonas fluorescens-based expression platform, Pfēnex Expression TechnologyTM, has been engineered to overcome the shortcomings of traditional expression platforms. This has been done by creating an off the shelf toolbox of expression plasmids and host strains. The elements of expression strategy, gene cloning, and host cell phenotypes combine seamlessly in a completely harmonized platform to deliver robust protein expression strains, using high throughput, parallel screening methods on more than a thousand unique strains in as little as four weeks.
3:50 Networking Refreshment Break
4:15 A Cell-Free Translation System for Producing Complex Disulfide Bonded Proteins at the Multiliter Scale
Christopher Murray, Ph.D., Senior Director, Research, Sutro Biopharma
Sutro’s OCFS system produces complex eukaryotic proteins such as cytokines and Fab antibody fragments up to 100 L scale at yields approaching 1 g/L. The open system allows us to model and rapidly optimize the system without the requirement of tuning more complex cellular networks required to maintain cell viability. The linear scalability of the system means that engineered proteins identified in high-throughput screens and selections can be immediately scaled for production of gram to kilogram quantities, thus avoiding the delays and challenges of conventional mammalian cell line development.
4:45 Harnessing Proteomics to Engineer a Better Protein Expression System
Stephen P. Chambers, Ph.D., Vice President, Protein Production, AbPro
We utilize proteomic-wide access to the whole of the human genome to assist in the selection and improvement of protein expression hosts. We will be rolling out the 3,000 proteins encoded by the secreted genome, to be followed by the 3,000 proteins of the druggable genome. This tool will allow protein researchers to more efficiently produce their proteins of interest.
5:15 Micro-Algae as a Platform for the Production of Therapeutic Proteins
Steve P. Mayfield, Ph.D., Associate Dean, Department of Cell Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute
5:45 Welcoming Reception in the Exhibit Hall
7:00 Close of Day
Links to Companion Meetings