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. 2017 Jun 9;9(6):945–958. doi: 10.1080/19420862.2017.1336592

Figure 3.

Figure 3.

Primary screening of humanized anti-GPVI Fabs. (A-B) Western blot analysis under non-reducing conditions of light chain expression using anti-human kappa chain as primary antibody (A) or Fd expression using anti-human IgG Fd region as primary antibody (B). Deposit of 10 μL of supernatant. Numbers refer to ACT0xx variants. (a) trastuzumab Fab transiently expressed in similar conditions. (b) abciximab (Reopro®) 0.5μg. (C-D) ELISA selection of the best GPVI binders after developing with anti-human IgG (Fab specific)-peroxidase conjugate (C) or PpL-Peroxidase conjugate (D). Culture supernatants were analyzed after successive dilutions (1:10, 1:100, 1:1000) in C or (1:10, 1:50, 1:100, 1:1000 in D). Orange bars (−): culture supernatant with no Fab.