The aim of the present study was to optimize the conditions of antibody arrays for detecting Hippocalcin-like 1 (HPCAL1), phosphatidylethanolamine binding protein 1 (PEBP1), lectin galactoside-binding soluble 7 (LGALS7), and serpin peptidase inhibitor clade E member 2 (SERPINE2) as biomarkers for pancreatic cancer detection in a single assay and to investigate antibodies' specificity and cross-reactivity. Here, LGALS7 is linked to familial pancreatic carcinoma.