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. 2023 Apr 18;11(7):962–977. doi: 10.1158/2326-6066.CIR-22-0705

Figure 6.

Figure 6. WTX-330 is inducible, stable in human serum, and selectively processed by dissociated primary human tumor samples. A, The components of WTX-330, including fully human IL-12 (yellow), a half-life extending HSA-specific single-domain antibody (blue), the activity blocking Ab domain (teal), and the protease-cleavable linkers (red). B, In vitro activity of WTX-330 in the HEK-Blue IL-12 reporter assay comparing intact (blue), and protease-activated (cleaved) WTX-330 (red) to rhIL-12 (black). C, In vitro activity of intact (blue) and cleaved (red) WTX-330 in primary human Tblasts compared with rhIL-12 (black). D, WTX-330 was diluted into healthy human serum from n = 6 donors and incubated at 37°C for 24 or 72 hours before WTX-330 processing was measured by capillary Western blot analysis. E, WTX-330 was exposed to primary human tumor samples (n = 88) or primary human healthy cells (n = 13) for 48 hours before INDUKINE protein cleavage was measured. For B and C, error bars represent the mean (± SD) of duplicate wells. For E, box plots represent the 25th and 75th percentile, whereas the line represents the median value for each indication.

WTX-330 is inducible, stable in human serum, and selectively processed by dissociated primary human tumor samples. A, The components of WTX-330, including fully human IL-12 (yellow), a half-life extending HSA-specific single-domain antibody (blue), the activity blocking Ab domain (teal), and the protease-cleavable linkers (red). B,In vitro activity of WTX-330 in the HEK-Blue IL-12 reporter assay comparing intact (blue), and protease-activated (cleaved) WTX-330 (red) to rhIL-12 (black). C,In vitro activity of intact (blue) and cleaved (red) WTX-330 in primary human Tblasts compared with rhIL-12 (black). D, WTX-330 was diluted into healthy human serum from n = 6 donors and incubated at 37°C for 24 or 72 hours before WTX-330 processing was measured by capillary Western blot analysis. E, WTX-330 was exposed to primary human tumor samples (n = 88) or primary human healthy cells (n = 13) for 48 hours before INDUKINE protein cleavage was measured. For B and C, error bars represent the mean (± SD) of duplicate wells. For E, box plots represent the 25th and 75th percentile, whereas the line represents the median value for each indication.