Countercurrent tangential chromatography for large-scale protein purification

Oleg Shinkazh, Dharmesh Kanani, Morgan Barth, Matthew Long, Daniar Hussain, Andrew L. Zydney

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Recent advances in cell culture technology have created significant pressure on the downstream purification process, leading to a "downstream bottleneck" in the production of recombinant therapeutic proteins for the treatment of cancer, genetic disorders, and cardiovascular disease. Countercurrent tangential chromatography overcomes many of the limitations of conventional column chromatography by having the resin (in the form of a slurry) flow through a series of static mixers and hollow fiber membrane modules. The buffers used in the binding, washing, and elution steps flow countercurrent to the resin, enabling high-resolution separations while reducing the amount of buffer needed for protein purification. The results obtained in this study provide the first experimental demonstration of the feasibility of using countercurrent tangential chromatography for the separation of a model protein mixture containing bovine serum albumin and myoglobin using a commercially available anion exchange resin. Batch uptake/desorption experiments were used in combination with critical flux data for the hollow fiber filters to design the countercurrent tangential chromatography system. A two-stage batch separation yielded the purified target protein at >99% purity with 94% recovery. The results clearly demonstrate the potential of using countercurrent tangential chromatography for the large-scale purification of therapeutic proteins. Biotechnol. Bioeng. 2011; 108:582-591.

Original languageEnglish (US)
Pages (from-to)582-591
Number of pages10
JournalBiotechnology and bioengineering
Volume108
Issue number3
DOIs
StatePublished - Mar 1 2011

Fingerprint

Countercurrent Distribution
Chromatography
Purification
Proteins
Inborn Genetic Diseases
Resins
Buffers
Anion Exchange Resins
Myoglobin
Bovine Serum Albumin
Column chromatography
Recombinant Proteins
Fibers
Cardiovascular Diseases
Cell culture
Washing
Cell Culture Techniques
Technology
Desorption
Ion exchange

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Shinkazh, Oleg ; Kanani, Dharmesh ; Barth, Morgan ; Long, Matthew ; Hussain, Daniar ; Zydney, Andrew L. / Countercurrent tangential chromatography for large-scale protein purification. In: Biotechnology and bioengineering. 2011 ; Vol. 108, No. 3. pp. 582-591.
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Countercurrent tangential chromatography for large-scale protein purification. / Shinkazh, Oleg; Kanani, Dharmesh; Barth, Morgan; Long, Matthew; Hussain, Daniar; Zydney, Andrew L.

In: Biotechnology and bioengineering, Vol. 108, No. 3, 01.03.2011, p. 582-591.

Research output: Contribution to journalArticle

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