Pancreatic cancer is currently the fourth most common internal cancer worldwide. With more than 200,000 reported cases annually, its mortality is almost equal to the incidence rate. Pancreatic cancer is an aggressive malignancy with one of the worst outcomes of all cancers and has a very poor prognosis: with a 1-year survival of less than 25%, and once the tumors have metastasized, a five year survival of less than 5%. In our study, we aim to investigate the key proteins from the primary tumor of pancreatic ductal adenocarcinoma (PDAC) that drives metastatic spread. Quantitative profiling of proteins using dimethyl labeling followed by 2D-LCMS/MS analysis of five primary tumours from patients with observed lymph node metastases (TNM classification=N1) were compared against five primary tumours from patients with no lymph node metastases (TNM classification=N0). We have identified and quantified a total of 2451 proteins in at least 3 of 5 patients, of which 105 proteins displayed altered levels in N0 versus N1 (p < 0.1, delta fold change >0.5). For selected proteins, the findings were validated by immunohistochemistry and Western blotting.  Our data revealed a number of proteins that had already been related to cancer metastasis and tumour cell invasion such as dermatopontin, biglycan, activated leukocyte cell adhesion molecule (ALCAM), but also other potential new prognostic biomarkers such as DNA binding protein A and guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-2. Furthermore, our data indicate a number of proteins which were related to immune responses and tumour microenvironment. Taken together, the preliminary findings from this study shed lights on novel candidates, which may play pivotal roles in cancer cell invasiveness and metastatic spread in pancreatic cancer.