One-Step Recovery of scFv Clones from High-Throughput Sequencing-Based Screening of Phage Display Libraries Challenged to Cells Expressing Native Claudin-1

Figure 1

Library screening and analysis of sequences. (a) The panel shows the DNA fragments, gel-purified from sublibraries, after the indicated selection cycles. The corresponding plasmid preparations were digested with NcoI and XhoI restriction endonucleases, to release the DNA fragments encoding for the VH regions of the scFv fragments. The fragments were bar-coded and subjected to high-throughput sequencing, as described in the text. SM, size marker. (b) The chart reports the entropy values for the populations of fragments originating from the indicated selection cycles, after sequencing. (c) The reported values indicate the total number of clones, and the relative representation of the most abundant clone, within the corresponding selection cycles. (d) The chart indicates the relative distribution of clones, according to the number of counts observed, within the indicated ranges, for each of the 4 selection cycles. Cycles 3 and 4 show similar distributions.

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