Affinity-Tagged Protein Purification With an Automated System Increases Access to Purified Recombinant Proteins
May 23, 2007
Dr. Laurie Usinger
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Abstract:
Affinity-based protein purification is an increasingly common method used for generating milligram quantities of target protein for use in functional and structural experiments. The Profinia protein purification system has been developed to automate affinity purification and desalting of tagged proteins. The system requires minimal user interaction and can purify target protein in quantities of 1100 mg in 30 minutes. The instrument uses optimized, preprogrammed methods and prefilled reagents and cartridges to provide fast, reproducible, and cost-effective protein purification. In this presentation, examples of both histidine- and GST-tagged protein purification will be shown to demonstrate the consistent results that can be obtained from using automated purification methods. In addition, hands-on labor time will be compared with traditional techniques, such as gravity flow columns. Improvements in the speed and automation of routine affinity purification can improve laboratory efficiency and the consistency of results across multiple users. The Profinia system increases the ease and reproducibility of affinity techniques so all members of a laboratory can produce quality target protein.
About the speakers:
Laurie Usinger, PhD, was the lead scientist for the Biotechnology Explorer program at Bio-Rad Laboratories from 1996 to 2001, and was instrumental in the launch of seven classroom kits in the first year of the program. The kits teach the basic workflows of molecular biology, from gene expression to protein purification, and use Green Fluorescent Protein (GFP), expressed from the pGLO plasmid, as the primary teaching tool. Dr. Usinger has since held the following positions at Bio-Rad:
- Lead scientist — Precision Protein standards project (under the guidance and management of Roger Provost), 1998–2000
- Project manager and lead scientist — Development and launch of Precision Plus Protein standards project, 2002
- Project manager — Launch of Precision Plus Kaleidoscope standards, 2004
- Program manager — Profinia protein purification system, 2004–2006
- Lead scientist — Construction and design of consumables line supporting the Profinia protein purification system and management and coordination of the consumables with the instrument platform, 2004–2006