Application of Agrocybe aegerita Lectin 2 for Detecting GlcNAcylated Plasma Proteins Inpatients with Breast Cancer
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Abstract:
The level of terminal β-N-acetylglucosamine (GlcNAc) - modified protein in plasma was detected using enzyme-linked immunosorbent assay (ELISA) with reverse Agrocybe aegerita lectin2 agglutinin (AAL2). The protein and GlcNAcylated protein levels of three candidate tumor biomarkers - plasma protease C1 inhibitor (C1Inh), serpin B4, and serotransferrin (TF) - in plasma were determined by western blotting and reverse AAL2-based ELISA. When compared to healthy volunteers (HE),in the breast cancer (BC) group, the protein levels of C1Inh and TF in plasma decreased significantly, whereas the level of serpin B4 protein increased significantly, and the level of GlcNAcylated protein showed the opposite trend (p<0.05). Additionally, between the HE group and early-stage BC samples, the area under the receiver operating characteristic (ROC) curve (AUC) for C1Inh, serpin B4, and TF proteins was 0.66, 0.62, and 0.73, respectively, whereas that for GlcNAcylated C1Inh, serpin B4, and TF was 0.69, 0.76, and 0.83, respectively. The combined detection of GlcNAcylated serpin B4 and TF showed improved performance for differentiating BC from HE, with the AUC for HE/early-stage BC samples, HE/limited-stage disease, and HE/extensive-stage disease increasing to 0.89, 0.89, and 0.94, respectively. This study provides a theoretical basis for the detection of GlcNAcylated protein in plasma using reverse AAL2-based ELISA, and GlcNAcylated serpinB4 and TF can serve as potential diagnostic markers of BC.
