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. 2020 Mar 9;12(1):1731938. doi: 10.1080/19420862.2020.1731938

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© 2020 Merck Healthcare KGaA. Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 7. — T-cell activation by reconstituted CD3xHer2 bsAbs. SK-BR-3, a Her2-expressing human breast cancer cell line, were used as antigen-presenting cells, and were incubated for 6 h with TCR/CD3 (NFAT) effector cells and reconstituted CD3xHer2 Fab (blue) or CD3xHer2 VHH (green) bsAbs in different concentrations ranging from 0.01 nM to 100 nM. CD3xHer2 binds simultaneously to TCR/CD3 on effector cells and antigen on tumor target cells and stimulates NFAT luciferase activity. BioGlo Luciferase substrate was added for luminescence readout. Luminescence signal was monitored and relative luminescence units (RLU) represent normalized luminescence to untreated cells. Reagents TCEP and DHAA used for bsAb reconstitution were added in different concentration to investigate potential assay interference. Cetuximab (orange): Negative control; monovalent anti-CD3 oaSEED (gray).