A Biparatopic Intrabody Renders Vero Cells Impervious to Ricin Intoxication.

Abstract

Expression of camelid-derived, single-domain antibodies (V H Hs) within the cytoplasm of mammalian cells as "intrabodies" has opened-up novel avenues for medical countermeasures against fast-acting biothreat agents. In this report, we describe a heterodimeric intrabody that renders Vero cells virtually impervious to ricin toxin (RT), a potent Category B ribosome-inactivating protein (RIP). The intrabody consists of two structurally defined V H Hs that target distinct epitopes on RT's enzymatic subunit (RTA): V9E1 targets RTA's P-stalk recruitment site, and V2A11 targets RTA's active site. Resistance to RT conferred by the biparatopic V H H construct far exceeded that of either of the V H Hs alone and effectively inhibited all measurable RT-induced cytotoxicty in vitro . We propose that targeted delivery of bispecific intrabodies to lung tissues may represent a novel means to shield the airways from the effects of inhalational RT exposure.