Higher Throughput Protein Purification
January 23-24, 2013
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THURSDAY, JANUARY 24
7:15am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee
7:45 Chairperson’s Remarks
William Gillette, Ph.D., Senior Scientist, Protein Expression Lab, SAIC-Frederick, Inc., Frederick National Laboratory for Cancer Research (FNLCR), NIH
7:50 Expressing the Human Proteome for Structural Genomics and Generation of Affinity Reagents
Susanne Gräslund, Ph.D., Principal Investigator, Biotechnology, Structural Genomics Consortium, University of Toronto- Biography
The Structural Genomics Consortium, founded in 2004, has to date deposited more than 1300 structures in PDB. The high-throughput methods used to generate the proteins required, in various expression hosts, will be presented. Since 2011, the SGC also drives a large-scale project to generate renewable recombinant antibodies to Epigenetics proteins. Specific for this project is the high-throughput production of in vivo biotinylated antigens to be used in phage-display selections and the production of Fabs and whole IgG molecules.
8:20 Accelerating Process Development Cycle by Online, Real-time Systems, HT Robotic Purification Platform and Chip-Based Assays
Richard Wu, Ph.D., Senior Scientist, Process & Product Development (P&PD), Amgen, Inc.- Biography
With the aims of process efficiency and cost saving in biopharmaceutical companies nowadays, real time manufacturing processes are being implemented more frequently for both upstream cell culture and bioreactor development, and downstream purification processes. Due to the fact that such manufacturing processes are a relatively new concept in the biopharmaceutical industry, obtaining baseline data to understand and develop the new process through design of experiment (DOE) will require advanced technologies to meet the analytical needs with accuracy, precision, and timely manner. For both upstream and downstream process development, automatic online/real-time process monitoring technologies will allow close, precise control monitoring, and on-the-spot mitigation of process performance should adverse events occur during the processes. Proposed Micro Sequential Injection (µSI) technique mixed with advanced high throughput, high resolution ultra-performance liquid chromatography (UPLC) enables the advancement of traditional amino acid assay, glycan profiling assay, and other wet chemistry-based assays to become useful online tools to closely examine and monitor process performance and product quality attributes in real-time.
8:50 Monolithic Columns for High-Throughput Chromatography of (Glyco-) Proteins: Column Formats and Modes of Separation
Michael R. Buchmeiser, Ph.D., Professor & Vice Dean, Institute of Polymer Chemistry, University of Stuttgart- Biography
This talk will address relevant aspects of both protein analysis and purification using polymer-based, monolithic columns and sorbents. Issues related to high-throughput analysis and processing and to the isolation of specific proteins, e.g., glycoproteins will be presented. Furthermore, the requirements for such separations in terms of structure of the polymeric supports and functional groups to be used will be outlined.
9:20 Versatile Toolbox for High-Throughput Biochemical and Functional Studies with Fluorescent Fusion Proteins
Garwin Pichler, Ph.D., Scientist, Ludwig Maximilians University, Munich- Biography
To fully characterize proteins, it is crucial to determine biochemical characteristics such as peptide binding, DNA binding, enzymatic activity and complex formation. Here, we demonstrate an easy, reliable and versatile medium/high-throughput method to study biochemical and functional characteristics of fluorescent fusion proteins. Using a new system based on 96-well micro plates comprising an immobilized GFP-binding protein (GFP-multiTrap), we performed fast and efficient one-step purification of different fluorescent fusion proteins from cellular lysates. We present an innovative approach to analyze binding specificities of fluorescently labeled fusion proteins and to detect endogenous interacting factors in a fast and reliable manner in vitro.
9:50 Accelerating Cell Line Optimization and Process Development Using Automated Solutions for Protein Purification and CharacterizationJeremy Lambert, Strategic Marketing, PerkinElmerLabChip® microfluidics technology enables multiple protein critical quality attributes. The new JANUS® Biotx Pro provides researchers with automated small-scale purification using a choice of batch binding in filter plates, tip-based chromatography, and robotic-controlled mini-columns to enable rapid “click and go” sample preparation resulting in shorter development cycles and increased efficiency.
10:05 Coffee Break in the Exhibit Hall with Poster Awards
10:45 MTPP™: Multi-Target Parallel Processing or Turning a Dozen New Targets into a SBDD Pipeline in Less Than a YearPeter Nollert, Ph.D., CTO, Emerald Bio
11:15 High-Throughput Screening in Purification Process Development
Yiming Yang, Senior Scientist, Purification Process Development, Shire Human Genetic Therapies - Biography
A high-throughput screening (HTS) platform with Tecan automated system and Atoll Mini-Columns was utilized in a Design of Experiments (DoE) study to define a design space for a downstream process. The HTS system was also used in chromatography resin/condition screening. The application of the HTS method greatly increased the efficiency, shortened the development time, saved the material consumption, and improved process understanding.
11:45 High-Throughput Screening Technologies as a Tool in Purification Development of Biotherapeutic Proteins
Bas Kokke, Ph.D., Project Leader, Downstream Processing, Synthon BV
We investigated a proper method to predict dynamics of a chromatography column from the experimental data obtained using 96-well microtiter plates. The approach is based on implementing the well-known batch uptake principle into microtiter plate geometry. The specific adsorption coefficient (ka) for a resin under certain conditions will be determined from the microtiter plate based batch experiment. Examples of quantitative estimations of dynamic binding capacities of a model protein on Protein A and Cation Exchange are given.
12:15 pm Close of Morning Session
12:30 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own
1:30 End of Conference
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