Day 1 | Day 2
Thursday, August 27, 2009
7:15am Breakfast Presentation Sponsored by 
One-Button, One-Step, Tag-Free Affinity Purifications
Dennis C. Yee, Ph.D., Research & Development Group Leader, Lab Separations Research & Development, Bio-Rad Laboratories
Two unique tools that increase the efficiency and effectiveness of affinity purifications are now available. The Profinity eXact tag is a novel affinity tag that utilizes a Subtilisin protease immobilized resin to achieve—in one elution step—tag-free, purified protein. The Profinia System offers simple, one-button programming to obtain purification and desalting of affinity-tagged proteins. This presentation will focus on:
• New expression and purification applications of the Profinity eXact tag
• The flexibility of the Profinia system in handling a range of affinity purification methods and tags
8:25 Chairperson’s Remarks
8:30 Novel Alternatives to Tag-based Affinity Purification
Jeffrey Culp, Ph.D., Associate Research Fellow, Primary Pharmacology Group, Research Centers of Emphasis, Pfizer Global Research and Development
The need to understand protein biology sometimes calls for expression and purification approaches designed to preserve protein function and avoid use of fused purification tags on the protein target. Examples are presented that use affinity capture through immobilized ligands, through target protein bound co-factors and indirect capture through an associated subunit.
9:00 Lore of the Tag: Affinity Purification of Proteases and Kinases
Alfredo Tomasselli, Ph.D., Research Fellow, Biophysical and Analytical Chemistry, Pfizer, Inc.
Examples will be covered regarding tagged proteases and kinases purified to homogeneity in highly active forms by using a combination of affinity tags, classical, and ligand specific affinity chromatography. Examples will be presented surrounding tag removal by autocleavage or by cleavage with specific proteases. Instances will be discussed where the tag was inserted, but not used because other purification strategies proved to be more efficient. Moreover, cases where the tag was not removed due to the fact that good enzymatic activity and high resolution crystal structures were achieved will be discussed. The yields, binding efficiency, optimization, speed, reproducibility and scalability of the purification process will be presented along with the in depth characterization of the final product to establish its authenticity and biochemical activity.
10:00 Networking Coffee Break with Exhibit and Poster Viewing
10:30 Optimizing Protein Purification at Micro Scale
William Gillette, Ph.D., Senior Scientist, Protein Purification Group Leader, Protein Expression Laboratory, Advanced Technology Program, SAIC-Frederick, Inc., National Cancer Institute at Frederick
For proteomic and core service laboratories charged with purifying numerous proteins, a method to screen the outcomes of different chromatographic approaches in parallel for multiple proteins/conditions would be extremely useful in reducing costs, increasing throughput and, more importantly, improving the success rate. We have incorporated a micro scale, parallel, protein purification platform which has proven effective in identifying optimal conditions and reducing the failure rate of purification. We routinely use the system for affinity tag optimization, determining binding efficiency, optimizing chromatography parameters and predicting purification yield. The results obtained from this platform, both quantitative and qualitative, have been predictive in 500-fold scale up experiments.
11:00 Grafting of Protein L-Binding Activity onto any Recombinant Antibody Fragments
Philippe Billiald, Ph.D., Professor, Biochemistry, University of Paris-South
We report a new universal system for the detection and purification of recombinantly expressed single-chain antibody fragments (scFvs). The system consists of grafting a peptide stretch of the V-KAPPA chain of an antibody that shows high affinity for Protein L from Peptostreptococcus magnus on to other single-chain antibodies (scFv) in case they have no affinity for Protein L, therefore transferring the binding property to Protein L.
11:30 Purification of Engineered, Retargeted Botulinum Neurotoxin Molecules as Protein Therapeutics
Matthew Beard, Ph.D., Senior Scientist, Molecular, Syntaxin Ltd.
By engineering recombinant botulinum neurotoxins to target distinct cell types Syntaxin Ltd. has created a new platform technology to generate protein therapeutics. Suitable for application in a wide range of diseases, our programs use affinity tagged proteins expressed in bacteria. We have optimized small scale, shake flask, systems to produce research molecules for discovery. We use a twin 5l fermentor system to optimize scale up before outsourcing our development scale protein production requirements. The presentation will discuss our core technology and how we have addressed the challenges of tag selection, molecule optimization, cleavage specificity, scalability and decision making about whether to reengineer and validate untagged molecules before development.
12:00pm End of The Bioprocessing Summit
For more information, please contact:
Margit Eder, Ph.D., Conference Director
Cambridge Healthtech Institute
Phone: 781-972-5478
E-mail: meder@healthtech.com
For sponsorship information, please contact:
Suzanne Carroll, Manager, Business Development
Cambridge Healthtech Institute
Phone: 781-972- 5452
E-mail: scarroll@healthtech.com