蛋白制剂制造会议 - Day 1

概要 | 短期研讨会 | Day 1 | Day 2

议程 (PDF : English)

Thursday, January 10

7:00am - 3:00pm Registration Open 

7:30 Morning Coffee

8:15 Chairperson乫s Opening Remarks

Opening Keynote Presentation

8:25 Overview of Protein Production Deb Chakravarti, Ph.D., Beckman Professor of Applied Life Sciences, Keck Graduate Institute.

Featured Presentation

9:05 Facilitating Membrane Protein Expression for Structural Studies Lawrence J. DeLucas, Ph.D., Professor and Director, The Center for Biophysical Sciences and Engineering, The University of Alabama at Birmingham The need for structural data on membrane associated drug targets is no secret. There are a number of future challenges for protein scientists. The first is the production of purified membrane proteins such as GPCRs for drug discovery applications. We have developed new lentiviral vectors (uniquely different than commercial) that provide a powerful capability for the expression of membrane as well as aqueous eukaryotic proteins. Expression and purification data will be presented from our multi year collaborative program researching the full-length Cystic Fibrosis Transmembrane Regulator Protein. In addition to the foundation research, data from industry-validated studies that are utilizing this expression system for the production of a wide range of high-value protein targets for structural analysis and subsequent drug development pipelines will be included.

9:45 Purification, Biochemical, Biophysical, Structural and Immunological Characterization of a Potential Vaccine Candidate
Indresh Srivastava, Ph.D., Associate Director, Vaccines Research, Novartis Vaccines & Diagnostics Inc.
Our focus is to develop effective vaccines against different diseases caused by pathogens such as HIV, HCV, Influenza, Men, Corona Virus and others.  We have successfully purified glycoproteins from HIV, HCV and SARS to homogeneity, and performed in-depth biochemical, biophysical, immunological, and structural characterizations of potential vaccine candidates using state-of-the-art analytical tools.  These well-characterized vaccines are evaluated in animal models to establish a correlation between structure and immunogenicity of potential vaccines.  Once the structural and conformational integrity of a potential vaccine is established, and after in vivo evaluation in animal models, an important step is to establish the stability and optimal formulation conditions of the vaccine.

10:15 Coffee Break in the Exhibit Hall

11:15 Purification of the Anti-Inflammatory Protein Tristetraprolin to Homogeneity 
Heping Cao, Ph.D., Research Biologist, Human Nutrition Research Center, USDA-ARS
Purification of active proteins to homogeneity is the pre-requirement for producing antibodies and pharmaceutical reagents, and for studying protein structure and function during the post-genomic era. Some proteins are difficult to be purified from any sources, probably due to insolubility, degradation, and low-level of expression. We developed a protocol for the purification of tristetraprolin (TTP), an anti-inflammatory protein with the potential as a therapeutic target for the prevention and/or treatment of inflammation-related diseases. The purified TTP was able to bind to tumor necrosis factor mRNA AU-rich sequence. Recombinant human and mouse TTP proteins were also successfully used for the production of high-titer antibodies and for the study of post-translational modifications.

11:45 Overcoming Challenges in Supplying Proteins for New Drug Discovery
Patrick Mollat, Ph.D., Head of Protein Sciences, Galapagos SASU
In the field of drug discovery, many different target proteins have to be prepared for various applications: enzyme kinetics, receptor binding studies, high-throughput screening, crystallization. Usually little information is available on expression and purification strategies of novel targets. Hence, many projects face the exciting and often unforeseeable challenge of obtaining the necessary amount of biologically relevant proteins against ambitious timelines. Achieving such versatility and speed requires constant adaptation of expression and purification strategies. This presentation will illustrate how we have managed these challenges in the cases of proteins such as the androgen receptor ligand binding domain, LDL related proteins and Dickkopf family proteins, exemplifying how success often is hidden in "the detail."

12:15pm Close of Morning Session

12:30 Luncheon Workshop r Lunch on Your Own 

1:45 Chairperson乫s Remarks

1:50 Zinc Finger Nucleases for Rapid and Efficient Gene Knock-out and Site-Specific Integration in Production Cell Lines
Philip D. Gregory, Ph.D., Vice President, Research, Sangamo BioSciences Inc.
Efficient site-specific addition or deletion of investigator-chosen DNA sequences remains experimentally intractable in the context of the mammalian cell nucleus despite its utility to protein production & biotechnology. Zinc finger nucleases (ZFNs) offer a general solution to this problem. We show here that the transient application of designed ZFNs is sufficient to promote the permanent disruption of targeted endogenous genes and efficiently generate novel 乬knock-out乭 cell lines with improved production characteristics. Moreover, such ZFNs also catalyze the site-specific integration of long DNA stretches into a predetermined native genomic location. This data highlights the unique outcomes enabled by ZFN-mediated genome engineering in host cell line optimization.

2:20 Applications of High Titre, Stable Mammalian Cell Lines in Preclinical Development
Andrew Sandford, Vice President, Selexis SA
For selection of the lead candidate protein, several variants must be rapidly produced. Within weeks of transfection we have identified clones with very high specific activities (>50 pcd) after screening no more than 50 clones. As no amplification is required, mutant cell lines are also not required. This allows us to use parental cell lines based on CHOK1 that grow to high cell density upon scale-up. Within 6 weeks of Transfection, we have obtained titers in shake flasks that exceed 2 gm/liter without any media optimization. We have used this approach to replace transient expression of preclinical lead proteins.

2:50 High Level Recombinant Protein Production: Beadle and Tatum in the 21st Century
W. Dorsey Stuart, Ph.D., President & Chief Executive Officer, Neugenesis Corporation
Filamentous fungi have been the object of academic and commercial interest for all of the 20th Century. Aspergillus is a workhorse producer of industrial enzymes. Neurospora crassa was used by Beadle and Tatum in the 1940s in their Nobel Prize winning work to help demonstrate that genes make enzymes. Neugenesis has utilized the enormous resource of academic research with Neurospora including the sequencing of the genome and a gene knockout project to engineer Neurospora host strains with the capability of producing recombinant biopharmaceuticals at useful commercial levels.

3:20 Technology Spotlight

3:35 Refreshment Break in the Exhibit Hall

4:30 Comparative Assessment of Yeast Platforms for the Production of Recombinant Proteins
Gerd Gellissen, Ph.D., Co-Founder & Chief Scientific Officer, PharmedArtis Gmb
Yeast provides attractive expression platforms that are distinguished by a growing track record as producers of valuable pharmaceutical and industrial proteins. However, all systems have drawbacks and limitations and it is evident that no single system is optimal for all proteins, and that it is advisable to assess several species in parallel for their capability to produce a particular protein. This presentation describes the production processes for some selected yeast-based products and provides insights into novel tools that enable the simultaneous assessment of a range of diverged yeast species with a single vector.

5:00 Production of Proteins in the Milk of Transgenic Goats
Harry Meade, Ph.D., Senior Vice President, R&D, GTC Biotherapeutics Inc.
This talk will first look at the process of producing proteins in the milk of transgenic goats. Then I will discuss ATryn, the first recombinant antithrombin approved for human use.

5:30 Production of Glyco-Optimized Antibodies with Enhanced Fc Function
Lynn F Dickey, Ph.D., Vice President, Research, Biolex Therapeutics
A glyco-optimized rituximab was expressed in the small aquatic plant Lemna. The optimized glycosylation was accomplished by co-expressing an interfering RNA (RNAi) construct targeting the endogenous alpha-1,3-fucoslytransferase (FucT) and beta-1,2-xylosyltransferase (XylT) genes (Cox et al., 2006). The resulting rituximab contained a single major G0 N-glycan without any detectable xylose or fucose with enhanced effector function.

6:00 Reception in the Exhibit Hall