Amber Codon Mutational Scanning and Bioorthogonal PEGylation for Mapping Antibody Binding Sites on Human Arginase-1
Jaime Fernández de Santaella,
Nikolaj G. Koch,
Lorenz Widmer
et al.
Abstract:Epitope mapping is crucial for understanding immunological responses to protein therapeutics. Here, we combined genetic code expansion and bacterial surface display to incorporate S-allylcysteine (SAC) into human arginase-1 (hArg1) viaMethanococcoides burtoniipyrrolysyl-tRNA synthetase. Using an amber codon deep mutational scanning and sequencing workflow, we mapped SAC incorporation efficiency across the hArg1 sequence, providing insights into structural and sequence dependencies of non-canonical amino acid i… Show more
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