Abstract: The invention relates to methods of producing at least one out-of-frame peptide of 5-40 amino acid residues by a cell, and to methods of identifying said at least one out-of-frame peptide. The invention further relates to the identified out-of-frame peptides, and epitopes and polyepitopes encompassing at least one of said out-of-frame peptides, and to a nucleic acid molecule encoding at least one of said out-of-frame peptides. The invention further relates to methods of inducing an immune response against at least one out-of-frame peptide, method of treating an individual suffering from a tumor, comprising providing said individual with at least one out-of-frame peptide, and to a pharmaceutical composition, comprising at least one out-of-frame peptide. The invention additionally relates to a kit for detecting expression of at least one out-of-frame peptide, and to methods of typing an individual for presence or absence of a cell that expresses at least one out-of-frame peptide.

Abstract: A method of identifying a neoantigen in a tumor of a subject is disclosed. The method comprises: (a) culturing cells of the tumor under conditions which generate single cell clones of the tumor; and (b) analyzing the single cell clones for expression of a neoantigen which is reactive with T cells from the subject, thereby identifying the neoantigen in a tumor of a subject.