[Abstract] Purpose: To screen and identify immunogenic membrane antigens associated with pancreatic cancer by applying proteomics techniques. Methods: The membrane proteins of pancreatic cancer cell line SW1990 were cultured, extracted and purified, and the membrane proteins were separated by two-dimensional bidirectional gel electrophoresis (2DE), and the parallel 2DE gels were stained with Komas Brilliant Blue and hybridized by immunoblotting, respectively. IgG was collected and purified from the sera of 66 clinical patients with pancreatic cancer and 24 patients with chronic pancreatitis, and immunoblot hybridization was performed with the membrane protein 2-DE parallel gels, respectively. Hybridization-positive protein sites were identified by applying matrix-assisted laser desorption ionization time-of-flight mass spectrometry with peptide mass fingerprint library. RT-PCR, real-time fluorescence quantitative PCR and Western blot were applied to verify the validity of the screened membrane antigens at both gene and protein levels at the cell line and tissue levels, respectively, and to compare the differences in gene and protein expression levels of the target membrane antigens in different pancreatic cancer cell lines and normal pancreatic tissues. Results A total of 9 positive spots were found in the immunoblot hybridization of the membrane protein of pancreatic cancer cell line SW1990 with serum IgG from pancreatic cancer patients, which were not duplicated with the 2 positive spots found in the IgG hybridization of chronic pancreatitis, and VDAC2 was identified as a very promising candidate membrane antigen by mass spectrometry analysis. As demonstrated by RT-PCR and Real time-PCR experiments, the gene of candidate membrane antigen VDAC2 was expressed in cells of pancreatic cancer cell lines SW1990, AsPc, and P3. Western blot results showed that the protein expression of candidate membrane antigen VDAC2 was significantly higher in pancreatic cancer cell lines than in normal pancreatic tissues. Conclusion: The membrane protein VDAC2 may be an immunogenic pancreatic cancer-associated membrane antigen, and its gene was expressed in pancreatic cancer cell lines SW1990, AsPc, and P3, and the protein expression level was significantly higher in pancreatic cancer cell lines than in normal pancreatic tissues.