Biopanning And Application Of Norfloxacin Mimotope And Construction Of Phage Display Library Target To Small Molecule

Norfloxacin（NFLX） is a kind of Fluoroquinolones，In this study, monoclonalantibody against NFLX was used as target for the biopanning of NFLX mimotopes byphage display technology, and developed Phage-ELISA and Phage-CLEIA based onNFLX mimotopes,and a phage display library was constructed for the mimotopebiopanning of small molecule.The monoclonal antibody against NFLX was used as target for biopanningNFLX mimotopes from PhD(C7C) and PhD(12).After being sequenced,12sequenceswere NKLRLPF、 TPSRLPF、 KFGFEPF、 TWKNLPF、 PIHRAPF、NTARWPF、 SPLRAPF、 NWSRLPF、 GPLYPGPFHTSH、ADWHQKFQMHSR、 SRMGPENWDKWY、 DHAYWPKFWGKA,the commonsequence of mimotopes from PhD(C7C) was XXXXXPF, in which X represented anyamino.Phage-ELISA was developed with clone H3, in which the IC50was56.1ng/mL,the linear working range was10.0~400.0ng/mL, the coefficients variation of meanintra-assay and inter-assay were5.7％and7.3％respectively. the recoveries of NFLXadded in feed(300、600、1000μg/kg) were varied between70.4％and120.3％.which variation coefficients was ranged from6.9%-9.3%.Phage-CLEIA was developed with clone H3, in which the IC50was41.7ng/mL,the linear working range was5.0~400.0ng/mL, the coefficients variation of meanintra-assay and inter-assay were7.2％and9.8％respectively, the recoveries of NFLXadded in feed(300、600、1000μg/kg) were varied between74.0％and118.0％.which variation coefficients was ranged from9.8%-11.2%.A phage display library (C7C) had been constructed,which complexity wasidentified to be2×107,almost fitted the need for biopanning.Otherwise, the aminoacid distribution of random display library was tested to be similarity to Theoreticalfrequency.