Display omitted •We for the first time develop a low-cost electrochemical immunosensor on paper for quantitative analysis of pancreatic cancer biomarker PEAK1.•The combination of electrochemical detection with gold nanoparticles enables high detection sensitivity.•Graphene oxide is drop-coated on carbon electrodes, providing a simple strategy to link antibodies to electrodes.•The immunosensor exhibits a low limit of detection of 10 pg mL−1 PEAK1, indicating high detection sensitivity for early cancer diagnosis.•This electrochemical platform has tremendous potential for point-of-care early diagnosis of a variety of cancers in resource-poor settings. Due to the lack of specific early detection methods for pancreatic cancer, it usually goes undetected until it is advanced. By employing paper-based electrodes (PPEs), herein we for the first time developed a disposable low-cost paper-based immunosensor for rapid early quantitative detection of pancreatic cancer with a new biomarker, pseudopodium-enriched atypical kinase one, SGK269 (PEAK1). The immunosensor was constructed by fabricating PPEs immobilized with the versatile nanomaterial graphene oxide for the incorporation of antibodies to form an immunosensing platform, without the need of complicated surface modification. After it was confirmed that the PPEs exhibited excellent electrochemical properties, a sandwich-type electrochemical immunosensor was subsequently constructed by employing graphene oxide layers immobilized with anti-PEAK1, and the antibody conjugated with gold nanoparticles (AuNPs-tagged-Anti PEAK1). Further, spectral and surface characteristic studies confirmed the formation of the immunosensing platform. The immunosensor for PEAK1 exhibited a wide linear range between 10 pg mL−1 and 106 pg mL−1 with a low limit of detection (LOD) of 10 pg mL−1. The obtained results point towards rapid, sensitive, and specific early diagnosis of pancreatic cancer at the point of care and other low-resource settings.