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. 2022 May 5;119(19):e2202439119. doi: 10.1073/pnas.2202439119

Fig. 1.

Fig. 1.

Major shed species of MSLN and mAb 15B6 binding to various forms of MSLN. (A and B) Cell culture medium from A431/H9 (A) or ascites from a mesothelioma patient (B) were purified on an SS1P antibody affinity column and shed MSLN sequence was determined by nano-LC-MS/MS as described in Materials and Methods. (C) Binding of various concentrations of mAb 15B6 to M and V variants of Fc-MSLN measured by ELISA. (D) Binding of Fab of mAb 15B6 to MSLN-His measured by biolayer interferometry (KD was calculated from individual concentrations). (E) Culture medium from OVCAR-8 cells (lane 1) and Fc-MSLN (lane 2) were analyzed by Western blot with either 15B6 or MN. (F) Binding of Fab of mAb 15B6 to MSLN–599-his but not to MSLN–585-his measured by biolayer interferometry. (G) ELISA measuring binding of mAb 15B6 to MSLN–599-his, MSLN–GPI, and MSLN–585-his.