| abstract |
Mutant epitopes encoded by oncogenes are almost always located inside cells, making them invisible to conventional antibodies. The inventors generated single chain variable fragments (scFv) specific for mutant peptides presented by human leukocyte antigen (HLA) molecules on the cell surface. These scFvs can be converted into full-length antibodies, called MANAbodies, targeting "mutation-associated neoantigens" that bind to HLA. Phage display libraries representing highly diverse arrays of single-chain variable region fragment sequences were first designed and constructed. Competitive selection methods are then used to identify clones specific for peptides that bind to predefined HLA classes. In this way, the inventors obtained scFvs, including one specific for peptides encoded by common KRAS mutants and another specific for peptides encoded by common EGFR mutants. Molecules targeting MANA can be developed that specifically react with mutant peptide-HLA complexes even when these peptides differ from the normal wild-type form by only one amino acid. |