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Affinity-Tagged Protein Purification With an Automated System Increases Access to Purified Recombinant Proteins
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Event Recorded on:
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Wednesday, 23 May 2007, 10:00 AM - 11:00 AM
(GMT -07:00) Pacific Time (US & Canada); Tijuana
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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 between 1–100 mg in 30 minutes. The instrument uses optimized, preprogrammed methods and prefilled reagents and cartridges to provide fast, reproducible, and cost-effective protein purification. Examples of both histidine- and GST-tagged protein purification will be presented 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 benefit laboratory efficiency and consistency of results across multiple users. The Profinia system improves the ease and reproducibility of affinity techniques so that all members of a laboratory can produce quality target protein.
About the speaker:
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Laurie Usinger
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Laurie Usinger, PhD, was the lead scientist for the Biotechnology Explorer program at Bio-Rad from 1996–2001, 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 utilize Green Fluorescent Protein (GFP), expressed from the pGLO™ plasmid, as the primary teaching tool. 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 managed the coordination of the consumables with the instrument platform, 2004–2006
Purification and preparation of fusion proteins and affinity peptides containing at least two adjacent histidine residues may require a license under US patent 5,284,933 and US patent 5,310,663, including foreign patents (assignee: Hoffmann-La Roche).
Expression and purification of GST fusion proteins may require a license under US patent 5,654,176 (assignee: Chemicon International).
*Note: This presentation utilizes internet audio. Please ensure that your computer system has audio capabilities before attending.
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