January 13 - 17, 2014
Renaissance Hotel and Palm Springs Convention Center Palm Springs, California

A Community Dedicated to the
Evolving Field and Future of Biotherapeutics
2014 Archived Content

SHORT COURSE 13: SAMPLE QC: AGGREGATION AND BEYOND 

 

Tuesday, January 14 | 5:00-8:00 pm

 

Buffer Optimization: Approaches and Applications
 

This course will focus on implementing a buffer “optimization” screen into a core service protein expression lab. We will look at assay development, how the assay dovetails with a microscale purification platform, how the results of the screen are interpreted and, ultimately, what real benefits are gained by the approach. In addition, we will compare methods for measuring proteins’ melting temperature and examine characterization of protein-ligand interactions

5:00 pm PART I 

BillGilletteWilliam Gillette, Ph.D., Senior Scientist, Protein Expression Lab, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research

 

I.   Why buffer optimization?
      a. What can we achieve? Case study from the literature
      b.  Is this useful for your protein?
II.  Choosing and implementing a buffer ‘optimization’ screen into a core service protein
      expression lab
      a.  Unique aspects
      b. Common aspects
      c. The assay: measuring a proteins’ melting temperature (Tm)
III.  Operational
      a.  How does the screen fit into the existing platforms
      b.  Interpreting data
      c.   Benefits
      d.  Drawbacks

6:30 Dinner Break

7:00  PART II 

AndyStephenAndrew Stephen, Ph.D., Senior Scientist, Protein Expression Lab, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research
 

I.    Measuring protein melting and aggregation
      a. Use of extrinsic dyes
      b. Use of intrinsic protein fluorescence
      c. Data analysis
II.  Case studies using thermal melting and aggregation analysis
      a. Protein characterization
      b. Buffer optimization
      c. Protein Ligand interactions
III. Lessons learned

8:00 Close of Short Course

Speaker Biographies

William Gillette, Ph.D., Senior Scientist, Protein Expression Lab, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research

Bill Gillette is currently focusing on high-throughput micro-scale purification as a means to identify positive constructs, optimization of chromatography and supporting analysis techniques to evaluate protein expression constructs and the success of the micro scale chromatography experiments. His work is in the context of the Protein Expression Laboratory that provides protein expression/purification core service to the laboratories of the NCI, NIH and USAMRIID. He received his Ph.D. in Microbiology from NC State University, Raleigh, NC.

Andrew Stephen, Ph.D., Senior Scientist, Protein Expression Lab, Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research

Andy Stephen received his Ph.D. in Biochemistry from the University of Liverpool, UK for purifying and characterizing a series of novel plant cysteine proteases. He did postdoctoral training at Washington University in St. Louis at the School of Medicine with Dr. Alan Schwartz on the role of the Ubiquitin activating enzyme in the degradation of proteins within the nucleus. At the UCLA Department of Biological Chemistry he worked with Dr. Lenny Rome characterizing vaults; large ribonucleoprotein particles with unknown function but whose expression is up-regulated in many drug resistant cancer cells. In 2000 he moved to the National Cancer Institute in Frederick, Maryland as a staff scientist in the Protein Chemistry Laboratory (PCL). The PCL provides supports to NCI investigators by using a variety of biophysical methods to characterize molecular interactions. The laboratory offers expertise in Surface Plasmon Resonance Spectroscopy and solution based fluorescence approaches. Andy is currently developing methods to understand how ligand binding can change the aggregation profile and thermal stability of proteins. 

 

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Training Seminars 

Mon-Tues, January 13-14 

Biologics Formulation and Delivery  

 


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