Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer mortality in the United States and the five year survival is only 1-5%. The best available treatment for pancreatic cancer patients is surgery which is only effective at early stages of disease. However, only about 15% of patients diagnosed with pancreatic cancer have early stage disease. Failure to diagnose pancreatic cancer at early stages and lack of effective systemic treatments has led to the high mortality seen in patients with this disease. Thus, better tools are needed for diagnosis and treatment of pancreatic cancer. As monoclonal antibodies have been effective in the treatment of other cancers we sought to develop antibodies with high specificity for pancreatic cancer. BALB/c mice were immunized with fresh pancreatic ductal adenocarcinoma samples or normal pancreatic tissue. Splenocytes from immunized mice were fused with SP2/0 Ag14 myeloma cells and hybridized cells were selected and cultured. The supernatants from the hybridized cell cultures were screened for cancer-reactive monoclonal antibodies on fixed frozen sections of pancreatic cancer, pancreatitis and normal pancreas. Cy-3 conjugated secondary antibody was used to detect section-bound monoclonal antibodies. Monoclonal antibodies, DHIC2 4-A10 and DHIC3 5-H10, were found to selectively detect antigens in specimens of pancreatic adenocarcinoma, liver cholangiocarcinoma, stomach adenocarcinoma, and lung adenocarcinoma. Flow cytometry using dispersed cells from the pancreatic cancer cell line, Panc1, revealed that these antibodies recognize cell surface antigens on these cells. The antibodies were also tested on normal tissues and were found to react with ductal cells in pancreas and liver. This technique for making tumor reactive monoclonal antibodies has the potential to identify reagents that may be useful in the diagnosis and treatment of this lethal disease.