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. 2011 Jul 21;6(7):e22533. doi: 10.1371/journal.pone.0022533

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Carlring et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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The presence of GL117-A20 bsAb (a) and 1C10-A20 bsAb (b) was verified by simultaneous recognition of anti-rat IgG and anti-mouse IgG in ELISA assay 1 as visualized by an increase in the OD reading at 450 nm compared to parental mAbs. Antigen recognition of β-gal and recognition by an anti-mouse IgG antibody was determined in ELISA assay 2 showing the presence of hybrid GL117-A20 bsAb after binding of the GL117 binding arm to β-gal and the A20 binding arm to anti-mouse IgG (c). CD40 specificity and subsequent recognition by an anti-mouse IgG antibody was determined in ELISA assay 3 and this showed the presence of 1C10-A20 bsAb by the binding of the 1C10-binding arm to CD40 and the A20 binding arm to anti-mouse IgG (d). Flow cytometric analysis on viable CD40L cells showed an increase in median fluorescence after incubation with 1C10-A20 bsAb but not GL117-A20 bsAb (e). Neg CTL = negative control.