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

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

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

Xin Ge,Daniel S C Yang,Kimberly Trabbic-Carlson,Bumjoon Kim,Ashutosh Chilkoti,Carlos D M Filipe,

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.

Self-cleavable stimulus responsive tags for protein purification without chromatography. Publishing Authors By Initials

X Ge,DS Yang,K Trabbic-Carlson,B Kim,A Chilkoti,CD Filipe,

For similar proteins: recombinant proteins: recombinant fusion proteins research abstracts see: proteins: recombinant proteins: recombinant fusion proteins research

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

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Journal of the American Chemical Society

VOLUME: 127

Page Numbers: 11228-9

Journal Abbreviation: J. Am. Chem. Soc.

ISSN: 0002-7863

DAY: 17

MONTH: Aug

YEAR: 2005

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

Number of References:

LANGUAGE: eng

NlmUniqueID: 7503056

Self-cleavable stimulus responsive tags for protein purification without chromatography. Keywords Mesh Terms:

KEYWORDS: Recombinant Fusion Proteins

MESH TERMS: isolation & purification

Chemical & Substance for Abstract: Self-cleavable stimulus responsive tags for protein purification without chromatography. Information

Substance Name: Elastin

Registry Number: 9007-58-3

Grant and Affiliation Information for Self-cleavable stimulus responsive tags for protein purification without chromatography.

AFFILIATION: Department of Chemical Engineering, McMaster University, 1280 Main Street West JHE-377, Hamilton, Ontario L8S 4L7, Canada.

Country: United States

AGENCY: United States PHS

GRANT: R01 610232

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MEDLINETA: J Am Chem Soc

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