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Escherichia coli skp chaperone coexpression improves solubility and phage display of single-chain antibody fragments.

Authors
  • Hayhurst, A
  • Harris, W J
Type
Published Article
Journal
Protein expression and purification
Publication Date
Apr 01, 1999
Volume
15
Issue
3
Pages
336–343
Identifiers
PMID: 10092493
Source
Medline
License
Unknown

Abstract

Expression of single-chain antibody fragments (scAb)in the periplasm of Escherichia coli often results in low soluble product yield and cell lysis. We have increased scAb solubility and prevented cell culture lysis by coexpressing the E. coli Skp chaperone gene. A mutant Skp cistron was linked to a bacteriophage T7 gene 10 translational initiation region and placed either downstream of a scAb gene within an isopropyl beta-d-thiogalactopyranoside-inducible expression cassette or on a separate colE1-compatible arabinose-inducible vector. Increases in scAb solubility reflected the amount of coexpressed Skp. A bacteriophage display vector that was also engineered to coexpress Skp permitted display of a virtually undisplayable scAb and should prove useful in expanding library sizes.

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