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2014 Archived Content

Cambridge Healthtech Institute’s Sixth Annual
Protein Purification and Recovery
Streamlining Processes
January 13-14, 2014


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TUESDAY, JANUARY 14

7:15 am Conference Registration

7:30 Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee


Purifying Kinases 

8:30 Chairperson’s Remarks

Vinodh Kurella, Ph.D., Research Fellow, Bioinformatics, Harvard Medical School

8:35 The Purification of JAK Kinases

Jiansheng Wu, Ph.D., Scientist, Protein Chemistry, Genentech, Inc.

JAK1, JAK2, JAK3 and TYK2 belong to the JAK family of kinases, which have important roles in immune systems. Specific kinase inhibitors to these kinases are being developed by many companies for different indications. A key hurdle to these programs is to purify these kinases in sufficient quantity and quality to support high-throughput screening and crystallography due to the poor chromatography behavior of these proteins. We developed a robust purification process to overcome these issues. We purified up to 8mg highly active proteins from 1 L BEVS cells consistently, and these proteins were successfully used to run HTS and SAR assays.

9:05 Expression, Purification and Characterization of Recombinant Necroptotic RIP Kinases

Alexei DegterevAlexei Degterev, Ph.D., Associate Professor, Biochemistry, Tufts University

Homologous Receptor Interacting Protein (RIP) kinases 1 and 3, components of the Tumor Necrosis Factor alpha receptor signaling complexes, have recently emerged as critical mediators of regulated necrotic cell death, termed necroptosis. In my talk, I will discuss expression and purification of catalytically active recombinant kinases using baculoviral system, assays to measure their activity and inhibitor interactions and recently described crystal structure and small molecule inhibitors of RIP1 kinase and cellular necroptosis.

9:35 Extended Q&A

9:50 Coffee Break in the Exhibit Hall With Poster Viewing


Purifying Membrane Proteins 

10:50 Exploring Affinity Tags for Expression, Purification and Recovery of G Protein-Coupled Cannabinoid Receptor Type II (CB2)

Silvia Locatelli-HoopsSilvia Locatelli-Hoops, Ph.D., Scientist, Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism (NIAAA), National Institutes of Health (NIH) 

Human cannabinoid receptor type II (CB2), a G protein-coupled receptor (GPCR), regulates inflammation pathways. We explored the suitability of the affinity tags haloalkane dehalogenase (HaloTag) and Rho-Tag (rhodopsin C-terminal nonapeptide) for expression of CB2 in E. coli and for efficient purification of functional receptor. The use of different strains, induction methods and constructs with alternating tag positions was explored. Methods for purification of the receptor by tandem affinity chromatography and surface immobilization were developed allowing further structural and functional studies.

Qinghai Zhang S11:20 New Chemical Tools for Stabilizing Membrane Proteins

Qinghai Zhang, Ph.D., Associate Professor, Integral Structural and Computational Biology, The Scripps Research Institute

We have designed beta-sheet peptides and steroid-based facial amphiphiles to stabilize integral membrane proteins. These novel amphiphiles are structurally distinct from classical detergents, and have enabled structural analysis of several membrane proteins by X-ray crystallography or electron microscopy. We will focus on the discussion of these chemical tools and respective applications in the structural dynamics studies of ATP-binding cassette transporters.

11:50  Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own

1:30-2:00pm Conference Registration



BUZZ Sessions png

2:00 BuzZ Session A (More Details >>)

3:00 Refreshment Break in the Exhibit Hall with Poster Awards

3:45 BuzZ Session B (More Details >>)

4:45 Close of Conference


4:30-5:00 Short Course Registration

5:00-8:00 Dinner Short Courses (SC8-SC14) More Details >> 




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