eIF3d is a protein biomarker which has a potential for the diagnosis of various cancers. Herein, a bio-platform was constructed for eIF3d sensing by using LSG and surface functionalization with anti eIF3d antibody via EDC/NHS chemistry. Following the surface modifications, XPS and several electrochemical methods were used. Difference in the signals were related to biomarker amounts between 75–500 ng/mL. LOD was calculated as 50.4 ng/mL. Selectivity of biosensor was tested by using of various interference molecules. EIF3d was also successfully detected in synthetic biological samples. Thus, to the best of our knowledge, this study is one of the rare studies on use of LSGs in immunosensor studies.