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Affinity gains by target-binding screening and NGS approaches.
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posted on 2015-06-05, 03:26 authored by Dongmei Hu, Siyi Hu, Wen Wan, Man Xu, Ruikai Du, Wei Zhao, Xiaolian Gao, Jing Liu, Haiyan Liu, Jiong HongThe scFv-Fc mutants were expressed in Expi293 cells to determine their apparent binding affinities to ErbB2 by screening ELISA. The fold affinity enhancement was calculated as the EC50 ratio of wild-type HuA21/mutant. The numbers of mutants from different CDR libraries are indicated.
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antibody affinity maturationSequencing Technologies Phage display technologyDeep sequencingEffective Optimizationlibrary sequence diversityscreening approachacid saturation mutagenesisPotent candidatesbinding propertiesIllumina platformAntibody affinityPhage Display IntegratedCDR sequencesenrichment landscapes4 x 106 DNA sequencesNWGNGS analysisnwcgene librariesnovel library generationphage libraries25.5 pMacid substitutionsperturbation mutagenesis strategynsgcandidate identification
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