Simple bioseparations using self-cleaving elastin-like polypeptide tags

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Abstract

We introduce a new method for the purification of recombinant proteins expressed in Escherichia coli using self-cleaving elastin-like polypeptide (ELP) fusion tags without the need for affinity chromatography or proteolytic tag removal. Using this method we obtained high purity, activity and reasonable yields for ten diverse target proteins.

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Figure 1: Purification of ELP-intein–tagged protein.
Figure 2: Purification of test proteins.

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Acknowledgements

Thanks to M. Hecht and L. Bradley from the Department of Chemistry at Princeton University for donating the gene encoding S-824. Thanks also to Y. Shi at the Department of Molecular Biology at Princeton University for his donation of and help with the AHSP protein. This work was partially supported by the US National Science Foundation Graduate Student Fellowship and US Army Research Office grant W911NF-04-1-0056.

Author information

Correspondence to David W Wood.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Building the ELP-intein expression construct. (PDF 35 kb)

Supplementary Fig. 2

Confirmation of full dissolution of precipitated ELP fusion proteins. (PDF 95 kb)

Supplementary Fig. 3

Purification of additional test proteins. (PDF 253 kb)

Supplementary Methods (PDF 159 kb)

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Banki, M., Feng, L. & Wood, D. Simple bioseparations using self-cleaving elastin-like polypeptide tags. Nat Methods 2, 659–662 (2005) doi:10.1038/nmeth787

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