Epitope binding analysis of human Fab by competition-ELISA.

<p>A and B, Fab anti-V m253 and m254 binding to the V-antigen were analysed by competitions with equal amount of control IgG1, mouse anti-<i>Burkholderia mallei</i> (Bm), and five other mouse anti-V mAbs (7.3 m, 10–1 m, 74–1 m, 84–1 m, 141–1 m) as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013047#s4" target="_blank">Materials and Methods</a>, respectively. The amount of bound human Fab was then determined. C. Lack of competition between human Fab anti-F1 fragment and mouse anti-F1(F1m) mAb. D. Competition among human antibodies. Fab m252, m253, and m254 were mixed with equal amounts of IgG m253 and applied to an ELISA plate coated with their respective antigens. The amounts of bound Fab were determined. PBS indicates m253 mixed with an equal volume of PBS buffer only. In all panels, NC indicates samples with no primary antibodies added serving as secondary antibody controls.