CONFERENCE SERIES: Bioprocess & Manufacturing
Recorded at: PEGS: The Essential Protein Engineering Summit  

Digital Course: Affinity Tags for Protein Purification

May 20, 2010

About this Product:
This digital course provides a great overview of affinity tags along with an excellent case study of developing tandem affinity purification (TAP)-mass spectrometry approaches to protein purification.  Bill Gillette (from SAIC-NCI Frederick) has personally purified over 800 unique proteins, and generously shares his extensive knowledge and experience including interactive discussions with the course participants.  Alexey Veraksa (from UMass Boston) discusses the advantages and pitfalls associated with the use of the original TAP tag and the more recently developed GS-TAP tag, and shares his group’s new data on developing even more efficient ways to isolate and analyze protein complexes.

Recorded at: 
Sixth Annual PEGS Summit, May 17-21, 2010, Boston, MA

About the Conference:
PEGS is the largest event on engineering of recombinant proteins and alternative scaffolds.
The 2010 event had over 1000 attendees, 9 cutting-edge programs, 68 exhibiting companies , 9 visionary Keynote presentations, and 100+ posters


Choosing the Right Tag: For You and Your Protein 
William Gillette William Gillette, Ph.D., Senior Scientist, Protein Expression Lab, SAIC-NCI Frederick
Affinity tags offer a wide array of features and drawbacks for both protein expression and purification. A discussion of the pros and cons of these tags will be followed by examples of how their use simplifies (or complicates) protein expression and purification in a Core Service lab.

Biography: The Protein Expression Laboratory provides protein expression/purification core service to the laboratories of the NCI, NIH and USAMRIID.  Dr. Gillette is currently focusing on micro-scale purification and supporting analysis techniques to evaluate protein expression constructs and the success of the micro-scale chromatography experiments.  Dr. Gillette earned his Doctorate in Microbiology from North Carolina State University, Raleigh, North Carolina.

Using Tandem Affinity Purification-Mass Spectrometry for Analyzing Intracellular Signaling Complexes
Alexey Veraksa Alexey Veraksa, Ph.D., Assistant Professor, Biology Department, University of Massachusetts, Boston
In this talk, I will use a case study of mapping the protein complexes in the Notch pathway to share our experiences with tandem affinity purification (TAP)-mass spectrometry approaches. The Notch signaling pathway is a conserved mechanism of intercellular communication that is used in all metazoans to establish molecular differences between adjacent cells. We have used the original TAP tag, as well as more recently developed GS-TAP tag, to analyze protein interactions in the Notch pathway. We have validated some of the novel interactions using functional assays in cultured cells and in vivo. I will discuss the advantages and pitfalls associated with the use of the TAP tags, and will share our new data on developing even more efficient ways to isolate and analyze protein complexes.

Biography: My laboratory studies signaling networks in Drosophila development, with a focus on dynamic protein-protein interactions that control developmental signaling pathways. As a postdoctoral fellow at the MGH Cancer Center, Harvard Medical School, I applied new protein purification methods in Drosophila and used them in combination with mass spectrometry to analyze protein complexes in the Notch interaction network. This study identified the Drosophila ß-arrestin Kurtz (Krz) as a Notch signaling modulator that regulates the turnover of the Notch receptor. After becoming an independent researcher at UMass Boston, I continue to investigate the diverse functions of ß-arrestin Krz in Drosophila development. We have recently found that Krz inhibits MAPK signaling at multiple developmental stages by a previously unknown mechanism. Part of our research efforts is devoted to developing new protein purification strategies that can be used for isolating and characterizing protein complexes involved in signal transduction. I received my Doctorate in Developmental Biology from the University of California, San Diego in La Jolla, California.

About this Product:
2 Presentations
Over 67 slides
150 minutes
Single Copy - $345.00
Site License - $1,380.00

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