Monoclonal antibodies (MAbs) reactive with Marburg virus (strain Musoke) were evaluated for both biological activity and specificity. Several of the Marburg virus- (MBGV) specific MAbs reduced the size and/or number of MBGV plaques in vitro. The ability of the MAbs to affect plaque formation in vitro was demonstrated to be specific for the glycoprotein (GP) of the strain of MBGV used for vaccination. Using deletion analysis and peptide mapping, the binding epitopes of several of these neutralizing MAbs were identified. Not unexpectedly, the epitopes were shown to lie in the most hypervariable and highly glycosylated region of MBGV GP. An analysis of the in vivo activity of several MAbs revealed that some antibodies provided substantial but incomplete protection of naive guinea pigs by passive transfer. These data suggest that neutralizing epitopes exist within MBGV GP but that induction of antibodies to these neutralizing epitopes may not be sufficient for protection from lethal infection.