Fig. 4

Exemplary ELISA analyses of constructed scFv-Fc antibody libraries. a-d The graphs represent the ELISA results of serial dilutions of 72 h time-point supernatants from CHOEBNALT85 cells transfected with 1 μg of HybriFree library pool DNA from the following captured cells: (a) chicken immunized with human DNase I; (b) mouse immunized with human artemin; and (c) rabbit immunized with GST-E2C protein. The irrelevant scFv-Fc vector was used as the negative control in each transfection. d ELISA results of single clone screening of the rabbit GST-E2C library pool shown in panel c. Wells G12 and H12 were transfected with library pool and negative control DNA, respectively. The clones gave highly positive OD signals ≥ 15 times over the negative control are highlighted in red. The clones with the signals 3–10 times higher than the negative control have orange highlighting. e Normalized artemin ELISA of 10 anti-artemin clones obtained from the immunized mouse (library pool ELISA results are shown in Fig. 3b). f Absorbance readouts of the same clones used in initial screening