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Purification of an elastin-like fusion protein by microfiltration.

Publication ,  Journal Article
Ge, X; Trabbic-Carlson, K; Chilkoti, A; Filipe, CDM
Published in: Biotechnology and bioengineering
October 2006

This article describes a simple and potentially scalable microfiltration method for purification of recombinant proteins. This method is based on the fact that when an elastin-like polypeptide (ELP) is fused to a target protein, the inverse phase transition behavior of the ELP tag is imparted to the fusion protein. Triggering the phase transition of a solution of the ELP fusion protein by an increase in temperature, or isothermally by an increase in salt concentration, results in the formation of micron-sized aggregates of the ELP fusion protein. In this article, it is shown that these aggregates are efficiently retained by a microfiltration membrane, while contaminating E. coli proteins passed through the membrane upon washing. Upon reversing the phase transition by flow of Milli-Q water, soluble, pure, and functionally active protein is eluted from the membrane. Proof-of principle of this approach was demonstrated by purifying a fusion of thioredoxin with ELP (Trx-ELP) with greater than 95% recovery of protein and with greater than 95% purity (as estimated from SDS-PAGE gels). The simplicity of this method is demonstrated for laboratory scale purification by purifying Trx-ELP from cell lysate using a syringe and a disposable microfiltration cartridge. The potential scalability of this purification as an automated, continuous industrial-scale process is also demonstrated using a continuous stirred cell equipped with a microfiltration membrane.

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Published In

Biotechnology and bioengineering

DOI

EISSN

1097-0290

ISSN

0006-3592

Publication Date

October 2006

Volume

95

Issue

3

Start / End Page

424 / 432

Related Subject Headings

  • Recombinant Fusion Proteins
  • Peptides
  • Industrial Microbiology
  • Filtration
  • Escherichia coli
  • Elastin
  • Biotechnology
  • Automation
 

Citation

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ICMJE
MLA
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Ge, X., Trabbic-Carlson, K., Chilkoti, A., & Filipe, C. D. M. (2006). Purification of an elastin-like fusion protein by microfiltration. Biotechnology and Bioengineering, 95(3), 424–432. https://doi.org/10.1002/bit.21046
Ge, Xin, Kimberly Trabbic-Carlson, Ashutosh Chilkoti, and Carlos D. M. Filipe. “Purification of an elastin-like fusion protein by microfiltration.Biotechnology and Bioengineering 95, no. 3 (October 2006): 424–32. https://doi.org/10.1002/bit.21046.
Ge X, Trabbic-Carlson K, Chilkoti A, Filipe CDM. Purification of an elastin-like fusion protein by microfiltration. Biotechnology and bioengineering. 2006 Oct;95(3):424–32.
Ge, Xin, et al. “Purification of an elastin-like fusion protein by microfiltration.Biotechnology and Bioengineering, vol. 95, no. 3, Oct. 2006, pp. 424–32. Epmc, doi:10.1002/bit.21046.
Ge X, Trabbic-Carlson K, Chilkoti A, Filipe CDM. Purification of an elastin-like fusion protein by microfiltration. Biotechnology and bioengineering. 2006 Oct;95(3):424–432.
Journal cover image

Published In

Biotechnology and bioengineering

DOI

EISSN

1097-0290

ISSN

0006-3592

Publication Date

October 2006

Volume

95

Issue

3

Start / End Page

424 / 432

Related Subject Headings

  • Recombinant Fusion Proteins
  • Peptides
  • Industrial Microbiology
  • Filtration
  • Escherichia coli
  • Elastin
  • Biotechnology
  • Automation