Selection of anti-zeranol ScFv antibody by phage display technology
ZHANG Qingqing
ZHANG Xiao
CHENG Luogen
LIU Yuan
TANG Min
LIU Xianjin
Abstract：Objective To screen the single chain antibody against zeranol(ZER)from the phage-display antibody library.Methods ELISA plate was coated by BSA-conjugated ZER,while subtractive panning was employed.The ratio of positive clone in the phage antibody library was elevated by three rounds of continual"adhesion-elution-amplification"solid panning,using two ways of eluting with trypsin and ZER.The positive clones was evaluated by phage ELISA.SDS-PAGE method was used to analyze the best time for getting more protein secreted form E.coli.Results Along with the increasing of elutriation rounds,specific phages were high enriched and 10 clones were screened from 6 pieces of 96 clones(OD value of positive clone is more 3 times than negative clone's).Five clones were transfected in HB2151 successfully and four different clones against ZER were found after sequenced.One of them was expressed prokaryotically,and the target protein possessed a protein's molecular weight of 28 KD.Conclusion This work provides a base for getting anti-ZER ScFv from human naive phage-display antibody library.
Keywords：phage-display antibody displaysingle chain antibodyzeranol
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 718-721 )