We compared further the amino acid sequences of AAL-2, GSL-II, WGA and HPA. Changes in glycosylation patterns play important functions in embryonic development, metabolic homoeostasis, pathogenic infections, immune surveillance and various diseases, including oncogenesis, diabetes and autoimmune disorders [1]. Therefore glycan analysis is usually of great importance for understanding the significance of glycan modifications. Lectins, with their diverse carbohydrate-binding properties, are potent tools for the study of glycan conjugates in various biological materials without glycan liberation. For example, lectins have been applied in the separation of cells at different stages, mitogenic activation of immune cells and identification of blood groups, as well FRP-1 as characterization of glycan alterations on the surface between normal and neoplastic cells [2C4]. The newly developed lectin array has been proved to be a powerful tool for the comprehensive analysis of glycans or glycoconjugates [5]. Much significant information has been obtained from use of lectin arrays. 1-2-Fucose-specific lectin (rBC2LCN) has been applied in the detection of undifferentiated induced pluripotent stem cells/embryonic stem cells, but not differentiated somatic cells [6], TKA (agglutinin) and PNA (peanut agglutinin) were used to distinguish the stem-like glioblastoma neurosphere cultured from a traditional adherent glioblastoma cell collection [7], GNA (agglutinin), NPA (agglutinin), PSA (agglutinin), LcH (lectin) and Con A (concanavalin A) have been utilized for the detection of high-mannose N-linked oligosaccharides on differentiated neutrophils in comparison with the promyelocytic leukaemia cell collection HL-60 [8], DSL (lectin), SLL [lectin; formerly LEL (lectin)] and MAL (lectin) showed a binding preference for mouse EW-7197 laminin, whereas SNA (agglutinin), SSA (agglutinin) and TJA-I (agglutinin-I) showed strong binding to bovine transferrin [9]. More lectins with unambiguous and/or unique glycan-binding selectivity are needed to be analyzed and exploited for probes of glycan structures. Thus there is an urgent need to find powerful lectin candidates for glycan analysis. Lectins are useful tools for tumour diagnosis, antivirus research and drug-delivery studies [10]. For example, MAL has been used in prostate malignancy diagnosis because of its preferential binding to prostate-specific antigen [11], HPA (agglutinin) recognizes the glycosylation changes of metastatic breast malignancy [12], BCA (lectin) can potently provide access inhibition of HIV-1 and influenza viruses [13], microvirin has anti-HIV-1 activity with a high security profile and low toxicity [14], and odorranalectin has been reported to be the smallest lectin so far and with potential for drug delivery and targeting [15]. An increasing quantity of lectins from plants and animals have been purified and characterized; however, the information on lectins isolated from fungal sources remains limited [16]. Fungal lectins are attractive because of their wide distribution, high content, varied carbohydrate-binding specificities and especially anti-tumour activities. lectin was shown to possess anti-tumour activity against human EW-7197 colon cancer HT29 and breast malignancy cell lines MCF-7 [17], lectin exhibited anti-proliferative activity in hepatoma HepG2 cells and human breast malignancy MCF-7 cells [18], lectin-1 and lectin-2 could inhibit the growth of sarcoma 180 cells [19], lectin exerted potent anti-tumour activity in mice bearing sarcoma 180 [20], and lectin was shown to be EW-7197 cytotoxic to HeLa cells [21]. In the present paper, we statement a novel fungal lectin AAL-2 (lectin 2) from your fruiting body of lectin-II), which have been widely used in biochemical and biomedical research. Moreover, we showed that AAL-2 experienced anti-tumour activity not only for the induction of hepatoma cells apoptosis was collected from your Sanming Institute of Fungi (Sanming, Fujian, P.R. China). GlcNAc was purchased from Sangon Biotech. Epoxy-activated Sepharose 6B was purchased from GE Healthcare. Cell lines and mice A murine hepatoma cell collection H22, and a human hepatoma cell collection Huh7, were provided by the CCTCC (China Center for Type?Culture Collection, Wuhan University or college). Male BALB/c mice (6C8-week-old) were purchased from your Hubei Experimental Animal Laboratory (Hubei, China) and managed in a pathogen-free facility. Procedures were performed according to approved protocols and in accordance with recommendations for the proper care and use of laboratory animals. Preparation of GlcNAc-coupled Sepharose 6B matrix GlcNAc-coupled Sepharose 6B was prepared according to the manufacturer’s instructions (GE Healthcare). In brief, 2?g of epoxy-activated Sepharose 6B was washed twice with 200?ml of distilled water, mixed with GlcNAc at a final concentration of 200?mol/ml in distilled water (pH?13) at a temperature.