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. Author manuscript; available in PMC: 2014 Feb 19.
Published in final edited form as: Circ Res. 2012 Mar;110(7):927–937. doi: 10.1161/CIRCRESAHA.112.265140

Figure 1. Functionality assessment of anti-VCAM1 nanobodies.

Figure 1

A and B: Flow cytometry analysis of anti-VCAM1 nanobodies on untreated VCAM1-negative (red) and TNFα-treated VCAM1-positive (blue) mouse bEND5 (A) or human HUVEC endothelial cells (B) (x: PE-A, log scale; y: %max). PE-labeled anti-VCAM1 monoclonal antibody was used as a positive control, whereas no nanobody and cAbBcII10 were used as negative controls. All 10 anti-VCAM1 nanobodies bound to mouse VCAM1-positive cells (A), and 6 out of 10 nanobodies were found to be crossreactive for human VCAM1-positive cells (B); C: Representative sensogram of cAbVCAM1-5 binding to mouse VCAM1. D: 99mTc-nanobodies bound to VCAM1-positive, TNFα-stimulated bEND5 cells. 99mTc-cAbVCAM1 binding to stimulated cells was significantly higher than binding to unstimulated cells, except for 99mTc-cAbVCAM1-8. Binding was successfully blocked by an excess of unlabeled nanobody, thereby demonstrating specificity. * P<0.05 vs 99mTc-cAbBcII10. † P<0.05 vs TNF (+) blocked condition.