Glucose is an important source of energy in the body, the primary substrate to provide energy of brain. Glucose monitoring in the brain need to be measured to comprehend glucose metabolism toward clinical purposes. Glucose sensors based on first generation design are widely used. In present study, manufacturing glucose sensors based on first generation type of sensors that consist of ruthenium-complexes coating and permselective membrane of poly (ophenylenediamine), with two immobilizing enzyme techniques is evaluated. Carbon fibre electrodes covered by ruthenium could enhance the sensitivity toward hydrogen peroxide up to three times higher compared to bare carbon fibre electrodes. The ruthenium deposition was reached optimum enhancement by applying constant potential at -500 mV vs. Ag/AgCl for 15 minutes in ruthenium solution pH 4.5. Perm-selective membrane was growth over ruthenium layer by electropolymerization process from monomer o-phenylenediamine at +700 mV vs. Ag/AgCl. The optimum performance of this membrane for hydrogen peroxide selectivity and ascorbic acid blockade was obtained by electropolymerization for 30 minutes. Enzyme layer was evaluated by comparing two different immobilization techniques; one-step deposition and separate deposition process. In this study, these techniques did not show glucose responses although in the previous work, the same responses were recorded for enzyme immobilization with one-step deposition. As a comparison, glucose sensors based on second generation fashion was constructed, the current response was positively correlated with concentration. The electrochemical process of glucose sensors construction has approved since the enhancement of hydrogen peroxide sensitivity and ascorbic acid blockade were performed. While, enzymatic aspect still has to be improved in order to provide enzyme layer as recognition and conversion site. However, further investigation for this process is still required to improve both processes.