Large-scale isolation of protein N-terminal peptides from LysargiNase digests was achieved by one-step strong cation exchange chromatography with disposable and parallel-processable pipette-tip columns. Novel cleavage sites for signal or transit peptides and non-canonical translation initiation sites were identified by the tip-based method. Together with two other methods similarly consisting of protease digestion and one-step tip LC, we have established a simple and comprehensive N- and C-terminomics approach called CHAMP (chromatographic amplification of protein terminal peptides).