Ribosome display is a complete in vitro system that permits selection of gene based on the physical association of its encoded message cognately to the protein it henceforth translates. The resulting ternary complexes of aptameric protein‐mRNA‐ribosome (ARM) specific for the ligand are immobilized on the designated solid phase ligand. Herein, green fluorescent protein (GFP) is fused to Cκ stuffer region as scaffold with 5′ T7 and Kozak/ATG sequences for transcription/translation‐coupled reactions. In non‐library modality, the presence of as low as 10 femtogram DNA rd4 construct of 1.2 kb, i.e., approximately 7,500 molecules, are sufficient for recursive rounds of selection by MAb anti‐GFP coated on solid phase. Next, ribosome display library constructed within site 6 and site 8/9 of GFP was employed for selecting aptameric protein that binds to human IgE on solid phase. An input 10 μg of 10‐mer aptameric GFP library for the respective dual site entails 7.5×1012 physical molecules (1.2 kb), expressed within a potential diversity of 1.0×1026 of possible peptide aptamers. Recombinant aptameric GFP was shown to bind to human IgE PS, BED, JW8, but not human IgG, lysozyme, BSA. Aptameric GFP also competes with the soluble receptor for binding human IgE on solid phase. Fragments of 25 to 100 bp from six IgE‐binding aptameric DNA by DNaseI were then shuffled, reassembled via internal homology by PCR. The molecularly evolved species exhibited improved binding to IgE. Thus, RD is an adequate technology platform for selecting aptameric GFP that specifically inhibits IgE binding to FcεRIα.
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