Hapten mediated display and pairing of recombinant antibodies accelerates assay assembly for biothreat countermeasures

Laura J. Sherwood, Andrew Hayhurst

    Research output: Contribution to journalArticlepeer-review

    15 Scopus citations

    Abstract

    A bottle-neck in recombinant antibody sandwich immunoassay development is pairing, demanding protein purification and modification to distinguish captor from tracer. We developed a simple pairing scheme using microliter amounts of E. coli osmotic shockates bearing site-specific biotinylated antibodies and demonstrated proof of principle with a single domain antibody (sdAb) that is both captor and tracer for polyvalent Marburgvirus nucleoprotein. The system could also host pairs of different sdAb specific for the 7 botulinum neurotoxin (BoNT) serotypes, enabling recognition of the cognate serotype. Inducible supE co-expression enabled sdAb populations to be propagated as either phage for more panning from repertoires or expressed as soluble sdAb for screening within a single host strain. When combined with streptavidin-g3p fusions, a novel transdisplay system was formulated to retrofit a semi-synthetic sdAb library which was mined for an anti-Ebolavirus sdAb which was immediately immunoassay ready, thereby speeding up the recombinant antibody discovery and utilization processes.

    Original languageEnglish (US)
    Article number807
    JournalScientific reports
    Volume2
    DOIs
    StatePublished - 2012

    ASJC Scopus subject areas

    • General

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