Figure EV1. In vitro characterization of the PSMA antibodies 10B3 vs J591.

• A
  Western blot analysis was performed using the indicated amounts of protein extracts prepared from 22Rv1^high cells or from PSMA‐negative HEK‐293 cells. Blots were analyzed using anti‐PSMA (mJ591 or m10B3) and anti‐tubulin as a loading control.
• B, C
  PSMA protein was immunoprecipitated from 22Rv1^high cells (B) and lung SCC samples (C) using chimeric J591, 10B3, and MOPC‐21 (control) antibodies. The inputs (5%) and immunoprecipitates (30%) were analyzed by Western blot using murine J591 antibody.
• D
  Mouse Sp2/0 cells transfected with human PSMA were incubated with the indicated concentrations of primary murine mAbs (mJ591, m10B3), followed by a chimeric J591 antibody (chJ591, 10µg/ml) and finally with fluorescence labeled mouse anti‐human antibody for specific detection of chJ591 as indicated.
• E, F
  Chimeric versions of both mAbs were bound to LNCaP cells (E) and to a protein A coated sensor chip (F) and analyzed by flow cytometry and SPR, respectively, as described in the Materials and Methods section.

Source data are available online for this figure.