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

Andrew Hayhurst, William J. Harris

Research output: Contribution to journalArticle

95 Citations (Scopus)

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 β-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.

Original languageEnglish (US)
Pages (from-to)336-343
Number of pages8
JournalProtein Expression and Purification
Volume15
Issue number3
DOIs
StatePublished - Apr 1999
Externally publishedYes

Fingerprint

Single-Chain Antibodies
Immunoglobulin Fragments
Bacteriophages
Solubility
Escherichia coli
Genes
Display devices
Thiogalactosides
Arabinose
Bacteriophage T7
Cell culture
Periplasm
Libraries
Cell Culture Techniques

Keywords

  • Chaperone
  • Dicistronic
  • Phage display
  • Single chain antibodies
  • Skp

ASJC Scopus subject areas

  • Biochemistry

Cite this

Escherichia coli Skp chaperone coexpression improves solubility and phage display of single-chain antibody fragments. / Hayhurst, Andrew; Harris, William J.

In: Protein Expression and Purification, Vol. 15, No. 3, 04.1999, p. 336-343.

Research output: Contribution to journalArticle

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