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Fig. 3 | Journal for ImmunoTherapy of Cancer

Fig. 3

From: Therapeutic efficacy of a novel humanized antibody-drug conjugate recognizing plexin-semaphorin-integrin domain in the RON receptor for targeted cancer therapy

Fig. 3

Schematic Structures of H5B14-based ADCs, drug conjugation profiles, and their plasma stability: a Schematic representation of H5B14-based ADCs. Both MMAE and duocarmycin [DCM] were conjugated to H5B14 by the valine-citruline dipeptide linker according to the manufacturer’s instruction (www.concortis.com). b HIC analysis of MMAE and DCM conjugated to H5B14. Individual H5B15-MMAEs or H5B14-DCMs with different numbers of MMAE or DCM [0–6] are marked as P0 to P6. The DAR combining P2, P4, and P6 was calculated at 3.76:1 for H5B14-MMAE and 3.73:1 for H5B14-DCM. c Dissociation MMAE or DCM from H5B14-based ADCs in human plasma. Both H5B14-MMAE and H5B14-DCM at 10 μg per ml were incubated with fresh human plasma at 37 °C for 10 days. The amount of free MMAE or DCM in plasma was determined using the LC-MS/MS method [35] with slight modifications [6]. Samples also were used for measuring MMAE or DCM conjugated H5B14 as detailed in Materials and Methods. A ratio from free MMAE to the total MMAE in H-Zt/g4-MMAE was calculated to determine the percentage of MMAE dissociated from H-Zt/g4-MMAE

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