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Self-cleavable stimulus responsive tags for protein purification without chromatography.

Publication ,  Journal Article
Ge, X; Yang, DSC; Trabbic-Carlson, K; Kim, B; Chilkoti, A; Filipe, CDM
Published in: Journal of the American Chemical Society
August 2005

A simple method to purify recombinant proteins is described by fusing a target protein with an intein and an elastin-like polypeptide that only requires NaCl, dithiothreitol, and a syringe filter to isolate the target protein from Escherichia coli lysate. This tripartite fusion system enables rapid isolation of the target protein without the need for affinity chromatography for purification or proteases for cleavage of the target protein from the fusion. The elastin-like polypeptide tag imparts reversible phase transition behavior to the tripartite fusion so that the fusion protein can be selectively aggregated in cell lysate by the addition of NaCl. The aggregates are isolated by microfiltration and resolubilized by reversal of the phase transition in low ionic strength buffer. After resolubilizing the fusion protein, the intein is activated to cleave the target protein from the elastin-intein tag, and the target protein is then isolated from the elastin-intein fusion by an additional phase transition cycle.

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Published In

Journal of the American Chemical Society

DOI

EISSN

1520-5126

ISSN

0002-7863

Publication Date

August 2005

Volume

127

Issue

32

Start / End Page

11228 / 11229

Related Subject Headings

  • Recombinant Fusion Proteins
  • Peptides
  • Microchemistry
  • General Chemistry
  • Filtration
  • Elastin
  • Chromatography
  • 40 Engineering
  • 34 Chemical sciences
  • 03 Chemical Sciences
 

Citation

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Ge, X., Yang, D. S. C., Trabbic-Carlson, K., Kim, B., Chilkoti, A., & Filipe, C. D. M. (2005). Self-cleavable stimulus responsive tags for protein purification without chromatography. Journal of the American Chemical Society, 127(32), 11228–11229. https://doi.org/10.1021/ja0531125
Ge, Xin, Daniel S. C. Yang, Kimberly Trabbic-Carlson, Bumjoon Kim, Ashutosh Chilkoti, and Carlos D. M. Filipe. “Self-cleavable stimulus responsive tags for protein purification without chromatography.Journal of the American Chemical Society 127, no. 32 (August 2005): 11228–29. https://doi.org/10.1021/ja0531125.
Ge X, Yang DSC, Trabbic-Carlson K, Kim B, Chilkoti A, Filipe CDM. Self-cleavable stimulus responsive tags for protein purification without chromatography. Journal of the American Chemical Society. 2005 Aug;127(32):11228–9.
Ge, Xin, et al. “Self-cleavable stimulus responsive tags for protein purification without chromatography.Journal of the American Chemical Society, vol. 127, no. 32, Aug. 2005, pp. 11228–29. Epmc, doi:10.1021/ja0531125.
Ge X, Yang DSC, Trabbic-Carlson K, Kim B, Chilkoti A, Filipe CDM. Self-cleavable stimulus responsive tags for protein purification without chromatography. Journal of the American Chemical Society. 2005 Aug;127(32):11228–11229.
Journal cover image

Published In

Journal of the American Chemical Society

DOI

EISSN

1520-5126

ISSN

0002-7863

Publication Date

August 2005

Volume

127

Issue

32

Start / End Page

11228 / 11229

Related Subject Headings

  • Recombinant Fusion Proteins
  • Peptides
  • Microchemistry
  • General Chemistry
  • Filtration
  • Elastin
  • Chromatography
  • 40 Engineering
  • 34 Chemical sciences
  • 03 Chemical Sciences