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

Andrew Hayhurst, William J. Harris

    Research output: Contribution to journalArticlepeer-review

    100 Scopus citations

    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

    Keywords

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

    ASJC Scopus subject areas

    • Biotechnology

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