Extended Data Fig. 10: Characterization of IL-4 and Neo-4 protease and serum stability.
From: Design of cell-type-specific hyperstable IL-4 mimetics via modular de novo scaffolds
a-f, Human or mouse IL-4 and Neo-4 were incubated with trypsin (a,b), chymotrypsin (c,d), or pepsin (e,f) for various time periods and subsequently subjected to SDS-PAGE analysis to detect protein degradation. Protease stability tests of each protein were performed once for each time point. g, 1 mg/ml (final concentration) of 6XHis-tagged hIL-4 or hNeo-4 was incubated in human serum for the indicated time periods up to 4 h. h, 1 mg/ml (final concentration) of 6XHis-tagged mIL-4 or mNeo-4 was incubated in mouse serum for the indicated time periods up to 4 h. Cytokines were detected with an anti-6XHis tag antibody via immunoblotting. hIL-4 and mIL-4 migrate as 22-24 kDa and 19-23 kDa, respectively, on the gel due to glycosylation. Serum stability test of each protein was performed once independently with similar results.
