Protein -- Pure And Simple

There are hundreds of ways to isolate recombinant proteins, many of which reliably yield large amounts of pure, fully functional product. Unfortunately, the most effective systems require the use of highly specialized chromatography reagents - for example, resins that have been chemically modified to assist in separation - making large-scale protein preparation an expensive prospect.

Now, David Wood and colleagues offer a remarkably simple alternative for the efficient preparation of fully functional proteins. The process relies on a polypeptide tag with keen sensitivity to temperature and salt levels; by modulating these conditions, the tag can be reversibly rendered either soluble or insoluble, allowing simple separation of the tagged protein from other cellular proteins by centrifugation. This tag is attached to the target protein by a self-cleaving linker, and once the tagged protein has been separated in initial rounds of centrifugation, the tag can be prompted to detach itself, leaving behind only the protein of interest.

The investigators test their technique with a wide range of proteins, and demonstrate that their process consistently generates highly purified proteins that retain biological function. According to the authors, this flexible and cost-effective system could potentially be employed for virtually any protein, and offers a level of simplicity that could benefit both small labs and large-scale proteomic research efforts.

Author contact:

David W. Wood, co-author (Princeton University, Princeton, NJ, USA)
Email: dwood@princeton.edu

Also available online.

(C) Nature Methods press release.

Message posted by: Trevor M. D'Souza