Autodisplayed Fv-antibody library for biomedical applications

In this work. the IgG VH chain Fv-library with randomized complementarity-determining region 3 (CDR3) region was expressed on the outer membrane of Escherichia coli using autodisplay technology. The target Fv-antibodies with binding activity to a target analyte were screened from the autodisplayed Fv-library on the E. coli outer membrane, and target clones were screened. Based on the binding properties of the screened Fv-antibodies, peptides with the screened clone of amino acid sequences of the CDR3 region were chemically synthesized and Fv-antibodies composed of CDRs and FRs was expressed as a fusion protein with green fluorescence protein (GFP). The binding properties of the synthetic peptides and Fv-antibodies with amino acid sequences of CDR3 regions from the selected clones were analyzed using fluorescence imaging and flow cytometry, and the affinity constants (Kd) of each peptide for binding to target antigens were calculated by fitting based on the isotherm model. The Fv-antibody library was applied for the screening of (1) inhibitor of monoamine oxidase B (MAO-B), (2) binding probes to monosodium urate (MSU) crystal, and (3) anti-Spike protein antibodies of SARS-CoV 2 and so on.

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