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• W2794223318 startingPage "e52" @default.
• W2794223318 abstract "Peptide-expressing phage display libraries are widely used for the interrogation of antibodies. Affinity selected peptides are then analyzed to discover epitope mimetics, or are subjected to computational algorithms for epitope prediction. A critical assumption for these applications is the random representation of amino acids in the initial naïve peptide library. In a previous study, we implemented next generation sequencing to evaluate a naïve library and discovered severe deviations from randomness in UAG codon over-representation as well as in high G phosphoramidite abundance causing amino acid distribution biases. In this study, we demonstrate that the UAG over-representation can be attributed to the burden imposed on the phage upon the assembly of the recombinant Protein 8 subunits. This was corrected by constructing the libraries using supE44-containing bacteria which suppress the UAG driven abortive termination. We also demonstrate that the overabundance of G stems from variant synthesis-efficiency and can be corrected using compensating oligonucleotide-mixtures calibrated by mass spectroscopy. Construction of libraries implementing these correctives results in markedly improved libraries that display random distribution of amino acids, thus ensuring that enriched peptides obtained in biopanning represent a genuine selection event, a fundamental assumption for phage display applications." @default.
• W2794223318 created "2018-03-29" @default.
• W2794223318 creator A5001523592 @default.
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• W2794223318 creator A5070736617 @default.
• W2794223318 creator A5083491181 @default.
• W2794223318 date "2018-02-06" @default.
• W2794223318 modified "2023-10-10" @default.
• W2794223318 title "Phage display peptide libraries: deviations from randomness and correctives" @default.
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• W2794223318 doi "https://doi.org/10.1093/nar/gky077" @default.
• W2794223318 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5961013" @default.
• W2794223318 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/29420788" @default.
• W2794223318 hasPublicationYear "2018" @default.
• W2794223318 type Work @default.
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