Disclosed are methods for identifying functional neoantigen peptides and using them in
cancer immunotherapy. Candidate HLA-restricted peptides are derived e.g., from tumor mutations, loaded onto paramagnetic nano-
artificial antigen-presenting cells (nano-aAPCs) that provide
peptide- HLA (
signal 1) and costimulatory ligands (
signal 2), incubated with T cells, magnetically enriched, expanded in culture, and characterized for
antigen specificity,
phenotype, and function to select therapeutic peptides. The
workflow supports
peptide cocktails across Class I / IIHLA presentation for heterozygous subjects, and a wide binding¬ affinity range. Resulting T-
cell products are enriched for central /
effector memory and polyfunctional responses. The methods enable vaccines, polymeric aAPC therapeutics, adoptive
cell therapies, and TCR- based biologies (e.g., bispecific T-
cell engagers).