Fig. 1

Screening and identification of anti-MUC18 mAb. Binding of JM1-24-3 to its target on melanoma cell surface is verified by FACS and ELISA a FACS analysis of JM1-24-3 binding on the surfaces of live melanoma cells showed that high metastatic cell lines A2058 and WM266-4 had significantly higher binding compared to low metastatic cell line A375. b ELISA analysis of JM1-24-3 binding with the immobilized melanoma cell lysates showed similar binding pattern as of FACS. High metastatic cell line A2058 showed the maximum binding. MUC18 protein is defined as the target of JM1-24-3 by immunoprecipitation (IP) and mass spectrometry (MS) analyses c Immunoprecipitation of WM266-4 lysates with JM1-24-3 showed two bands - 135kD (high intensity) and 110kD (faint band) by SDS-PAGE Coomassie blue staining. d MS analysis showed that MUC18 glycoprotein had invariably the highest score among all the hits for both the bands