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. 2008 Nov 10;105(46):17670–17675. doi: 10.1073/pnas.0805130105

Fig. 4.

Fig. 4.

DEL4000 library selection with polyclonal human IgG. (A) Plot representing the frequency (i.e., sequence counts) of DEL4000 library members after selection on polyclonal human IgG resin, as revealed by high-throughput 454 sequencing. The chemical structures of some of the most relevant compounds enriched are indicated. The building blocks used in the 2 synthetic steps are indicated in green and red, respectively, together with an identification number. (B) Affinity chromatography of CHO cells supernatant on resin containing the compound 02-40, spiked with human IgG labeled with either Cy5 or biotin. For antibody purifications, relevant fractions were analyzed by SDS/PAGE both with Coomassie blue staining and with a specific detection method (Cy5 fluorescence and a streptavidin horseradish peroxidase-based blot, respectively). M, molecular weight marker; In, input fraction for the chromatographic process; W, pooled washed fractions; E, pooled eluted fractions. The lane (+) corresponds to Cy-5- or biotin-labeled polyclonal human IgG. In, W, and E fractions were normalized to the same volume before SDS/PAGE analysis. The SDS/PAGE gels containing the fractions from the purification of both Cy5- and biotin-labeled IgG were analyzed by conventional staining with Coomassie blue, or by using a label-specific procedure (fluorescence imaging for Cy5-labeled IgG or streptavidin-based blot for biotinylated IgG; see also SI Text).