Gaze estimation, as a technique that reflects individual attention, can be used for disability assistance and assisting physicians in diagnosing diseases such as autism spectrum disorder (ASD), Parkinson’s disease, and attention deficit hyperactivity disorder (ADHD). Various techniques have been proposed for gaze estimation and achieved high resolution. Among these approaches, electrooculography (EOG)-based gaze estimation, as an economical and effective method, offers a promising solution for practical applications. Objective: In this paper, we systematically investigated the possible EOG electrode locations which are spatially distributed around the orbital cavity. Afterward, quantities of informative features to characterize physiological information of eye movement from the temporal-spectral domain are extracted from the seven differential channels. Methods and procedures: To select the optimum channels and relevant features, and eliminate irrelevant information, a heuristical search algorithm (i.e., forward stepwise strategy) is applied. Subsequently, a comparative analysis of the impacts of electrode placement and feature contributions on gaze estimation is evaluated via 6 classic models with 18 subjects. Results: Experimental results showed that the promising performance was achieved both in the Mean Absolute Error (MAE) and Root Mean Square Error (RMSE) within a wide gaze that ranges from −50° to +50°. The MAE and RMSE can be improved to 2.80° and 3.74° ultimately, while only using 10 features extracted from 2 channels. Compared with the prevailing EOG-based techniques, the performance improvement of MAE and RMSE range from 0.70° to 5.48° and 0.66° to 5.42°, respectively. Conclusion: We proposed a robust EOG-based gaze estimation approach by systematically investigating the optimal channel/feature combination. The experimental results indicated not only the superiority of the proposed approach but also its potential for clinical application. Clinical and translational impact statement: Accurate gaze estimation is a key step for assisting disabilities and accurate diagnosis of various diseases including ASD, Parkinson’s disease, and ADHD. The proposed approach can accurately estimate the points of gaze via EOG signals, and thus has the potential for various related medical applications.