An approach for N-linked glycan identification from MS/MS spectra by target-decoy strategy

Abstract Glycan structure determination serves as an essential step for the thorough investigation of the structure and function of protein. Currently, appropriate sample preparation followed by tandem mass spectrometry has emerged as the dominant technique for the characterization of glycans and glycopeptides. Although extensive efforts have been made to the development of computational approaches for the automated interpretation of glycopeptide spectra, the previously appeared methods lack a reasonable quality control strategy for the statistical validation of reported results. In this manuscript, we introduced a novel method that constructed a decoy glycan database based on the glycan structures in the target database, and searched the experimental spectra against both the target and decoy databases to find the best matched glycans. Specifically, a two-layer scoring scheme for calculating a normalized matching score is applied in the search procedure which enables the unbiased ranking of the matched glycans. Experimental analysis showed that our proposed method can report more structures with high confidence compared with previous approaches.