This paper presents a novel framework for unobtrusive biometric authentication based on the spatiotemporal analysis of human activities. Initially, the subject’s actions that are recorded by a stereoscopic camera, are detected utilizing motion history images. Then, two novel unobtrusive biometric traits are proposed, namely the static anthropometric profile that accurately encodes the inter-subject variability with respect to human body dimensions, while the activity related trait that is based on dynamic motion trajectories encodes the behavioral inter-subject variability for performing a specific action. Subsequently, score level fusion is performed via support vector machines. Finally, an ergonomics-based quality indicator is introduced for the evaluation of the authentication potential for a specific trial. Experimental validation on data from two different datasets, illustrates the significant biometric authentication potential of the proposed framework in realistic scenarios, whereby the user is unobtrusively observed, while the use of the static anthropometric profile is seen to significantly improve performance with respect to state-of-the-art approaches.