Human Anti-ERBB2 Recombinant Antibody (clone B1D2) (CAT#: HPAB-1133-FY)

Figure 1 High Her2 expression is required for efficient targeted exosome uptake.

Parental HEK293, and HEK-Her2 overexpressing cell lines were incubated with exosomes as described above, followed by staining with WGA-Alexafluor555 (red) and Hoechst (blue) post-fixation. Images were acquired on the Opera high-throughput confocal microscope.

Longatti, A., Schindler, C., Collinson, A., Jenkinson, L., Matthews, C., Fitzpatrick, L., ... & Tigue, N. (2018). High affinity single-chain variable fragments are specific and versatile targeting motifs for extracellular vesicles. Nanoscale, 10(29), 14230-14244.

Figure 2 High affinity anti-Her2 targeted exosomes are required for efficient uptake into Her2 positive tumour cell lines.

The indicated cell lines were incubated with targeted (G98A, ML39 and B1D2) and irrelevant scFv CFSE-exosomes in duplicate for 4 h, and cells were analysed for CFSE-exosome uptake by flow cytometry.

Longatti, A., Schindler, C., Collinson, A., Jenkinson, L., Matthews, C., Fitzpatrick, L., ... & Tigue, N. (2018). High affinity single-chain variable fragments are specific and versatile targeting motifs for extracellular vesicles. Nanoscale, 10(29), 14230-14244.

Figure 3 Non-targeted and anti-Her2-scFv-exosomes can be generated via genetic manipulation of HEK293 cells.

Binding of recombinant Her2-Fc to immobilised exosomes by ELISA; irrelevant scFv (open circles), G98A scFv (squares), ML39 scFv (triangles) and B1D2 (closed circles). Error bars represent the SEM of three independent experiments where samples were analysed in duplicate.

Longatti, A., Schindler, C., Collinson, A., Jenkinson, L., Matthews, C., Fitzpatrick, L., ... & Tigue, N. (2018). High affinity single-chain variable fragments are specific and versatile targeting motifs for extracellular vesicles. Nanoscale, 10(29), 14230-14244.

Figure 4 Two-color immunofluorescence of anti-ErbB2 receptor-grafted peripheral blood T cells.

Nontransduced peripheral blood T cells and T cells grafted with anti-ErbB2 immunoreceptors with different affinities of the binding domain (no. 1–5) were simultaneously incubated with a PE-conjugated anti-CD3 mAb and an FITC-conjugated anti-human IgG1 Ab and analyzed by flow cytometry.

Chmielewski, M., Hombach, A., Heuser, C., Adams, G. P., & Abken, H. (2004). T cell activation by antibody-like immunoreceptors: increase in affinity of the single-chain fragment domain above threshold does not increase T cell activation against antigen-positive target cells but decreases selectivity. The Journal of Immunology, 173(12), 7647-7653.

Figure 5 Activation of anti-ErbB2 receptor-grafted T cells by solid phase bound Ag.

Serial dilutions of recombinant ErbB2-Fc fusion protein (A) or of an anti-human IgG-Fc Ab (B) were coated onto microtiter plates. Peripheral blood T cells were grafted with anti-ErbB2 immunoreceptors with different Kd values of the anti-ErbB2 scFv domain and incubated for 48 h (1 × 10⁴ receptor-grafted T cells/well). IFN-γ in the supernatant was recorded by ELISA.

Chmielewski, M., Hombach, A., Heuser, C., Adams, G. P., & Abken, H. (2004). T cell activation by antibody-like immunoreceptors: increase in affinity of the single-chain fragment domain above threshold does not increase T cell activation against antigen-positive target cells but decreases selectivity. The Journal of Immunology, 173(12), 7647-7653.