Rapid assembly of sensitive antigen-capture assays for Marburg virus, using in vitro selection of llama single-domain antibodies, at biosafety level 4

Laura J. Sherwood, Lisa E. Osborn, Ricardo Carrion, Jean L. Patterson, Andrew Hayhurst

    Research output: Contribution to journalArticlepeer-review

    47 Scopus citations

    Abstract

    There is a pressing need for rapid and reliable approaches to the delivery of sensitive yet rugged diagnostic assays specific for emerging viruses, to hasten containment of outbreaks when and wherever they occur. Within 3 weeks, we delivered an antigen-capture assay for Marburg virus (MARV) that was based on llama single-domain antibodies (sdAbs) selected at biosafety level 4. Four unique sdAbs were capable of independently detecting MARV variants Musoke, Ravn, and Angola without cross-reactivity with the 4 Ebola virus species. The unoptimized assays could be performed in <30 min and, at best, provided a visual read of 10-100 pfu in a 100-μL sample when a colorimetric substrate was used and 0.1-1 pfu when a chemiluminescent substrate was used. All the sdAbs were specific for nucleoprotein, with an assay sensitivity that was reliant on detergent-mediated exposure of polyvalent antigen. Our strategy highlights the potential of direct antibody selection on filoviruses as a guide for effective and fast diagnostic development.

    Original languageEnglish (US)
    Pages (from-to)S213-S219
    JournalJournal of Infectious Diseases
    Volume196
    Issue numberSUPPL. 2
    DOIs
    StatePublished - Nov 15 2007

    ASJC Scopus subject areas

    • Immunology and Allergy
    • Infectious Diseases

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