Fig. 5

High-throughput FLIP applied to the screening of IP-grade mAbs. a 46 mAb produced and purified by CDI laboratories were tested by FLIP and immunoblotting (Additional file 1: Figure S3). The FLIP signal (mAb FLIP subtracted from the control IgG FLIP) and the % of the immunoprecipitated target protein measured after immunoblotting (% tot. lys.) are reported together with the name of the target proteins and the code specific for each antibody. Negative FLIP values (for which the FLIP signal of the background IP is greater than the FLIP signal after IP with the considered mAb) are reported as zero. b The FLIP values calculated using ImageJ and the % tot. lysate index obtained after immunoblotting are plotted and the R² of the best fit trend-line is reported. The inset shows the plotted values for mAbs that had a positive FLIP value but did not show a band for the immunoprecipitated target protein by western analysis (FLIP false positives in yellow), mAbs that were shown not to be IP-competent by both FLIP and IP/western (in green), and mAbs that did not have a positive FLIP value but showed a band for the immunoprecipitated target protein by immunoblot analysis (FLIP false negatives in red). In blue are reported all the mAbs that show good accordance between FLIP and IP/western analysis