Alanine scan mutagenesis in CDR3 of 125-2HPublic
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The murine antibody, 125-2H, is a potent neutralizer of interleukin (IL-18), which is associated with inflammation. Because of its murine origins it is not a clinical candidate, but its IL-18 neutralizing potency is the standard to which human monoclonal antibodies to IL-18 are held. To determine the amino acid residues most essential to its interaction with IL-18, alanine scan mutagenesis was performed on the third complementarity-determining regions (CDR3) of the light and heavy chains. Following testing in a KG-1 assay for IL-18 neutralizing ability, it was determined that the alanine substitutions at four CDR3 amino acid residues of the heavy chain are most essential for IL-18 binding.
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