Figure 2.

High throughput generation and characterization of recombinant antibodies. (a) For each target antigen, 94 clones were screened in ELISA and sequenced to identify unique clones. The plot shows the number of antigens selected (x-axis) versus the number of unique positive antibodies generated (y-axis) for each antigen. Separate plots are presented for antigens produced in either bacterial or mammalian expression systems, illustrating the improved success rate for mammalian antigens. (b) Example specificity data for antibodies selected against Slam f9 (produced in the HEK293 mammalian expression system). All antibodies are screened against target antigen, the relevant fusion partner that was used in selection, keyhole limpet hemocyanin (KLH), thyroglobulin, myoglobin, cytochrome c, human IgG, laminin, fibronectin, α-glycerol phosphate dehydrogenase, and total protein lysates from zebra fish (D. rerio) and yeast (S. pombe). Results are shown for 22 different antibodies as well as our routine anti-desmin control (des-D7) and a no antibody control. Detection was via time resolved fluorescence and values are shown on a logarithmic scale. (c) Global summary of secondary ELISA data for all antibodies in secondary screening. Signal generated on specific antigen is shown for all 4,437 samples (solid block). Signal achieved on one of the ten irrelevant antigens (cytochrome c) is also shown.