Development and characterization of monoclonal antibodies against pancreatic cancer marker hippocalcin-like 1 protein.

Pancreatic cancer remains a major challenge for research studies and clinical management. Monoclonal antibodies (MAbs) against pancreatic cancer biomarkers could be used in the early diagnosis of pancreatic cancer. The hippocalcin-like 1 protein (HPCAL1) is expressed in most pancreatic cancer cell lines. The goal of this work was to generate anti-HPCAL1 MAbs that have potential clinical applications. Recombinant HPCAL1 protein was successfully produced in an Escherichia coli expression system and was used as immunogen in mice. MAbs against HPCAL1 were generated using the standard hybridoma technique. Anti-HPCAL1 MAb 1E10 detects denatured HPCAL1 protein in Western blot analysis and immunoprecipitates native HPCAL1 protein from cell lysates. MAb 5A5 detects HPCAL1 in formalin-fixed, paraffin-embedded pancreatic cancer cell lines, making it a potentially useful reagent for pathologists. The MAb pair 1E10 and 5A5 had the optimal binding affinity for soluble HPCAL1 detection in a sandwich ELISA system, which is a potential clinical detection method for pancreatic cancer. In conclusion, multi-functional anti-HPCAL1 MAbs 1E10 and 5A5 have been generated and characterized, which may be used in early diagnosis of pancreatic cancer.