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Non-chromatographic purification of recombinant elastin-like polypeptides and their fusions with peptides and proteins from Escherichia coli.

Publication ,  Journal Article
MacEwan, SR; Hassouneh, W; Chilkoti, A
Published in: Journal of visualized experiments : JoVE
January 1, 2014

Elastin-like polypeptides are repetitive biopolymers that exhibit a lower critical solution temperature phase transition behavior, existing as soluble unimers below a characteristic transition temperature and aggregating into micron-scale coacervates above their transition temperature. The design of elastin-like polypeptides at the genetic level permits precise control of their sequence and length, which dictates their thermal properties. Elastin-like polypeptides are used in a variety of applications including biosensing, tissue engineering, and drug delivery, where the transition temperature and biopolymer architecture of the ELP can be tuned for the specific application of interest. Furthermore, the lower critical solution temperature phase transition behavior of elastin-like polypeptides allows their purification by their thermal response, such that their selective coacervation and resolubilization allows the removal of both soluble and insoluble contaminants following expression in Escherichia coli. This approach can be used for the purification of elastin-like polypeptides alone or as a purification tool for peptide or protein fusions where recombinant peptides or proteins genetically appended to elastin-like polypeptide tags can be purified without chromatography. This protocol describes the purification of elastin-like polypeptides and their peptide or protein fusions and discusses basic characterization techniques to assess the thermal behavior of pure elastin-like polypeptide products.

Duke Scholars

Published In

Journal of visualized experiments : JoVE

EISSN

1940-087X

Publication Date

January 1, 2014

Issue

88

Related Subject Headings

  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology
 

Citation

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MacEwan, S. R., Hassouneh, W., & Chilkoti, A. (2014). Non-chromatographic purification of recombinant elastin-like polypeptides and their fusions with peptides and proteins from Escherichia coli. Journal of Visualized Experiments : JoVE, (88).
MacEwan, S. R., W. Hassouneh, and A. Chilkoti. “Non-chromatographic purification of recombinant elastin-like polypeptides and their fusions with peptides and proteins from Escherichia coli.Journal of Visualized Experiments : JoVE, no. 88 (January 1, 2014).
MacEwan SR, Hassouneh W, Chilkoti A. Non-chromatographic purification of recombinant elastin-like polypeptides and their fusions with peptides and proteins from Escherichia coli. Journal of visualized experiments : JoVE. 2014 Jan 1;(88).
MacEwan SR, Hassouneh W, Chilkoti A. Non-chromatographic purification of recombinant elastin-like polypeptides and their fusions with peptides and proteins from Escherichia coli. Journal of visualized experiments : JoVE. 2014 Jan 1;(88).

Published In

Journal of visualized experiments : JoVE

EISSN

1940-087X

Publication Date

January 1, 2014

Issue

88

Related Subject Headings

  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology