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Impact of genetic engineering on downstream processing of proteins produced in E. coli

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Applied Molecular Genetics

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 43))

Abstract

Genetic engineering can be used to give a protein properties that are advantageous for downstream processing. Many heterologous proteins are degraded at high rates by proteases. Depending on which type of proteolytic degradation is encountered the strategy may be different: induction of inclusion bodies, change of the amino acid sequence in the sensitive site of the product, or protection by fusion of the product with other proteins. The number of unit operations needed to purify a protein may be reduced by addition of other polypeptides or amino acids to the product. Affinity chromatography, immobilized metal ion affinity chromatography, and extraction in aqueous two-phase systems are unit operations which can be made more versatile by the fusion technique.

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Abbreviations

Ala:

Alanine

Arg:

Arginine

Asn:

Asparagine

Asp:

Aspartic acid

DHFR:

Dihydrofolate reductase

E. coli:

Escherichia coli

Gly:

Glycine

His:

Histidine

IFN:

Interferon

IGF:

Human insulin-like growth factor

IgG:

Immunoglobulin G

IL:

Interleukin

kDa:

1000 dalton

NTA:

nitilotriacetate

ompT:

outer membrane protein T

PEG:

poly (ethylene) glycol

rDNA:

recombinant DNA

Ser:

Serine

SpA:

Staphylococcal protein A

ZZ:

artifical IgG-binding protein derived from staphylococcal protein A

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Authors and Affiliations

  1. Department of Biochemistry and Biotechnology, The Royal Institute of Technology, S-100 44, Stockholm, Sweden

    S. -O. Enfors, H. Hellebust, K. Köhler, L. Strandberg & A. Veide

Authors
  1. S. -O. Enfors
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  2. H. Hellebust
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  3. K. Köhler
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  4. L. Strandberg
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  5. A. Veide
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© 1990 Springer-Verlag

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Enfors, S.O., Hellebust, H., Köhler, K., Strandberg, L., Veide, A. (1990). Impact of genetic engineering on downstream processing of proteins produced in E. coli . In: Applied Molecular Genetics. Advances in Biochemical Engineering/Biotechnology, vol 43. Springer, Berlin, Heidelberg. https://doi.org/10.1007/BFb0009078

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  • DOI: https://doi.org/10.1007/BFb0009078

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  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-540-52794-7

  • Online ISBN: 978-3-540-47151-6

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