A human T cell epitope was mapped on tetanus toxin by the use of a simple method. Tetanus toxin peptide preparations were produced by various specific chemical and proteolytic cleavages. T cell proliferative response to these peptide preparations was assayed to determine which amino acid residues were present or absent in the T cell epitope. Cyanogen bromide-derived tetanus toxin peptides were separated by electrophoresis and transferred onto nitrocellulose. T cell proliferative response to nitrocellulose fragments was assayed to determine the molecular weight of the antigenic tetanus toxin peptide. The combination of the two procedures allowed us to define a 21-mer tetanus toxin peptide recognized by the T cell clone. This method represents an easily performed alternative for the mapping of T cell epitopes on protein antigen.