5C), providing the best separation (i.e., minimal false-negative and false-positive results). We correctly identified 27 of 30 patients with CC (90.0% sensitivity) and excluded 29 of 38 patients with benign duct disease (76.3% specificity).

For comparison, a conventional format of antibody-antibody sandwich system, i.e., MY.1E12-MY.1E12, was also performed. However, none of Temozolomide the obtained scores was better than those of the present WFA-MY.1E12 system: (P = 0.0138). The sensitivity was 56.7%; specificity, 84.2%; and AUC, 0.75 at a cutoff value of S/N = 9.36 (Fig. 5B and broken line in Fig. 5C). These results are better than those produced by biliary cytology (Table 3). Early and correct diagnosis of CC is still an urgent issue even with the aid of modern detection technologies. Although many CC-associated serological markers, such as CA19-9,6, 8, 9 MUC5AC,33 and Mac-2–binding protein,7 have been proposed, none of them has satisfactory sensitivity. With special focus on cancer-associated glyco-alteration, we recently proposed a robust strategy to develop high performance glycoprotein biomarkers using advanced technologies of glycopropteomics.17, 34 Along with the established strategy, we attempted differential glycan analysis using tissue sections containing both normal BDE and ICC Adriamycin chemical structure lesions from the same patients. An ultrasensitive glycan profiling technology, lectin microarray mined WFA30 as the most promising probe

to differentiate ICC from normal BDE with a significant P value (<0.0001). Subsequent histochemical analysis of 150 ICC sections using biotinylated WFA could confirm the strong expression of WFA-reactive glycans specific in cancerous lesions. The main aim of this study is to develop a robust diagnostic system targeting molecular bio-markers involved in body fluids. We have chosen bile as the primary target, because

the bile is in direct downstream of CC, and thus, is expected to contain much higher amounts of candidate marker molecules than in serum. In this study, we hypothesized that sialylated MUC1, established as an antigen for MY.1E12 monoclonal antibody and known to express well in BDE and CC cells, is one of the glycoproteins that carries the WFA-reactive glycans. As expected, staining with both MY.1E12 and WFA merged well in the cancerous Flucloronide lesions in ICC tissue sections (Fig. 3). In addition, the presence of sialylated MUC1 carrying WFA-reactive glycans was confirmed by western blot analysis using WFA-enriched bile fractions (Fig. 4). Therefore, it is conclusive that sialylated MUC1 is a carrier protein of WFA-reactive glycans in both ICC tissues and bile fluids. Thus, we developed a novel sandwich ELISA system that combined WFA and MY.1E12 to target bile samples (30 cases of CC and 38 cases of benign bile duct diseases). The sensitivity (90.0%) and AUC (0.86) obtained were superior to those produced by previous methods. Biliary cytology gave only poor sensitivity (10/18; 55.