In this commentary, we discuss seldom-noticed methodological problems affecting biological research on burnout and depression and make recommendations to overcome the limitations of past studies conducted in this area. First, we suggest that identified subtypes of depression (e.g., depression with melancholic features and depression with atypical features) should be taken into account in future biological research on burnout and depression, given that different subtypes of depression have been associated with distinct autonomic and neuroendocrine profiles. Second, we underline that research on burnout-depression overlap is made difficult by the absence of a consensual conceptualization and operationalization of burnout. In order to resolve this problem, we draw researchers’ attention to the urgency of establishing a commonly shared, clinically valid diagnosis for burnout. Finally, we question the possibility of identifying a biological signature for burnout in light of global research on burnout-depression overlap.