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. 2015 Oct 10;7(10):4067–4082. doi: 10.3390/toxins7104067

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© 2015 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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In vitro activity of dDTEGF13: (A) Bispecific DTEGF13 and its monospecific counterparts were tested and compared for their reactivity against MiaPaCa-2 cells. Proliferation assays were performed by analyzing ³H-thymidine uptake after a 72-h incubation with targeted toxins. Data are reported as percent control response. Each data point represents an average of triplicate measures ± SD; (B) Deimmunized DTEGF13 and non-mutated parental DTEGF13 were tested and compared for activity against HT-29 colon; and (C) PC-3 prostate carcinoma cells in thymidine uptake assays. Two batches are used for reproducibility; (D) A blocking assay was performed in which MiaPaca-2 pancreatic cancer cell lines were incubated with an inhibitory dose of dDTEGF13 and then blocked with increasing concentration of EGF13 ligand devoid of toxin. Thymidine uptake was then measured. The non-specific recombinant α-Ly5.2 was included as a negative blocking control.